Research ArticleClinical Studies

CD133(+)HIF-1α(−) Expression After Chemoradiotherapy Predicts Poor Prognosis in Rectal Cancer

YUICHI TACHIKAWA, KAZUSHIGE KAWAI, KOSUKE OZAKI, HIROAKI NOZAWA, KAZUHITO SASAKI, KOJI MURONO, SHIGENOBU EMOTO, JUNKO KISHIKAWA, YUICHIRO YOKOYAMA, SHINYA ABE, YUZO NAGAI, HIROYUKI ANZAI, HIROFUMI SONODA and SOICHIRO ISHIHARA
Anticancer Research April 2022, 42 (4) 2033-2043; DOI: https://doi.org/10.21873/anticanres.15684

Abstract

Background/Aim: CD133 and hypoxia-inducible factor 1α (HIF-1α) have been reported to be affected by chemoradiotherapy (CRT), but the combinatorial assessment of these markers for prognosis after CRT has not been fully investigated. Therefore, we aimed to predict recurrence and prognosis in patients with rectal cancer by assessing changes in the expression of both CD133 and HIF-1α after CRT. Materials and Methods: CD133 and HIF-1α expression was evaluated by immunohistochemistry in surgical specimens from 243 patients with advanced low rectal cancer who received CRT followed by curative resection. Results: The positivity rate of CD133 expression showed increase with increased HIF-1α expression. The combination of these two markers showed that the CD133(+)HIF-1α(−) group exhibited a markedly shorter relapse-free survival (p=0.007), higher liver recurrence (p=0.004), and higher local recurrence (p=0.006). Conclusion: CD133(+)HIF-1α(−) expression after CRT is a promising marker to predict recurrence in rectal cancer.

Key Words:

The current standard treatment for locally advanced rectal cancer is preoperative chemoradiotherapy (CRT) followed by total mesorectal excision (1, 2). Patients who receive CRT experience large variations in efficacy, and the prognoses of patients who undergo curative resection after CRT vary widely. We aimed to predict recurrence and prognosis in patients with rectal cancer by assessing the expression of key proteins in the tumor after CRT.

In the present study, we focused on the expression of two proteins after CRT, namely CD133 and hypoxia-inducible factor 1α (HIF-1α), both of which have been reported to be affected by CRT. CD133 is a tumor stem cell marker of colon cancer (3, 4). Some studies using colon cancer cell lines indicated that radiation therapy increased the protein and mRNA expression of CD133 (5, 6). High CD133 expression was reported to be associated with poor tumor regression grade after CRT in rectal cancer (7–11), but the reported prognostic impact of CD133 expression after CRT has been controversial. Some studies have indicated no correlation between high CD133 expression and prognosis (8), while others have indicated that high CD133 expression is associated with poor prognosis (10). Meanwhile, HIF-1α is a protein expressed under hypoxia, and high HIF-1α expression has been linked to tumor invasion, metastasis, and resistance to apoptosis, radiation, and chemotherapy (12, 13). Although it was reported that HIF-1α was up-regulated following exposure to radiation in endometrial, neck squamous, and colorectal cancer cell lines (14–16), HIF-1α expression after CRT has been reported to have no correlation with tumor regression grade (17, 18). Thus, the prognostic impact of post-CRT expression of HIF-1α also remains controversial (18, 19), similar to CD133.

CD133 and HIF-1α have been reported to exhibit co-regulated expression. Knockdown of CD133 decreased the expression of HIF-1α under hypoxic conditions in pancreatic cancer cell lines (20), and, conversely, knockdown of HIF-1α reduced the expression of CD133 in glioma cell lines (21). Furthermore, we previously reported that CD133(+) cells had higher HIF-1α expression during hypoxia than CD133(−) cells (22). However, the interaction between these two molecules has not been considered in most studies investigating the impacts of either CD133 or HIF-1α on prognosis after CRT. Therefore, in the present study, we assessed the expression of CD133 and HIF-1α in rectal cancer after CRT and investigated the correlation between these proteins and the recurrence of disease and prognosis.

Materials and Methods

Patients and tissue specimens. Two hundred eighty-four consecutive patients underwent curative resection after CRT for rectal adenocarcinoma at the University of Tokyo Hospital from September 2003 to December 2018. All patients were diagnosed with low rectal cancer, and the tumor depth was estimated to be deeper than the muscularis propria. Clinical diagnoses of all patients were stage I–IIIC rectal cancer based on the Union for International Cancer Control TNM (UICC TNM) classification (23). Patients received a total radiation dose of 50.4 Gy (1.8 Gy × 28 fractions) and concomitant 5-fluorouracil-based chemotherapy. Total mesorectal excision with lymph node dissection was performed 6–10 weeks after CRT. All patients underwent a standardized follow-up schedule that included assessment of carcinoembryonic antigen level every 3 months, chest-to-pelvic computed tomography every 6 months, and a colonoscopy every 12 months. Clinicopathological features were analyzed based on the UICC TNM classification (23) and World Health Organization histological criteria (24). An experienced pathologist evaluated the tumor specimens to determine the pathologic response to CRT according to the Japanese Classification of Colorectal, Appendiceal, and Anal Carcinoma (25). The pathologic response was graded as follows: grade 0, no regression; grade 1a, minimal effect (necrosis of less than one-third of the lesion); grade 1b, mild effect (necrosis of between one-third and two-thirds of the lesion); grade 2, moderate effect (necrosis of more than two-thirds of the lesion); and grade 3, no viable tumor cells (pathological complete response). Grade 3 was excluded because our immunohistochemical staining targeted residual cancer tissue. Finally, 243 patients were enrolled in the study. The study was approved by the ethics committee of the University of Tokyo [No. 3252-(10)]. The requirement for informed consent was replaced by the provision of information to the participants and the right of participants to opt out because of the retrospective nature of the study.

CD133 and HIF-1α immunohistochemical staining. Consecutive 4-μm sections fixed in formalin and embedded in paraffin were immunohistochemically stained using the technique described below. The tissues were treated with xylene and ethanol and then washed with phosphate-buffered saline (PBS). Endogenous peroxidase was blocked with 0.3% H2O2 in methanol for 20 min. After washing with PBS, heat-induced antigen retrieval was performed in ethylenediaminetetraacetic acid buffer (pH 8.0). Following incubation with a mouse monoclonal anti-CD133 antibody (dilution, 1:100; cat. no. 130-090-422; Miltenyi Biotec, Auburn, CA, USA) and mouse monoclonal anti-HIF-1α antibody (dilution, 1:300; cat. no. NB100-123; Novus Biologicals, Centennial, CO, USA), the Histofine SAB-PO (M) kit (Nichirei Corp, Chuo-ku, Tokyo, Japan) was used to prevent non-specific binding, treat with secondary antibody, and amplify the signal. For chromogenic development, the slides were incubated in 2% 3,3’-diaminobenzidine tetrahydrochloride and 50 mM Tris buffer (pH 7.6) containing 0.03% H2O2. Meyer’s hematoxylin (Sigma-Aldrich; Merck KGaA, St. Louis, MO, USA) was used for counterstaining for 30 s at 35-40°C.

Evaluation of CD133 and HIF-1α immunohistochemical staining. CD133 expression was defined as positive when CD133 staining was found in more than 5% of the entire tumor according to the method by Maeda et al., as described previously (26–28) (Figure 1A and B). Briefly, the slides were examined under a microscope at low power (from 40× to 200×) to identify the region containing the highest percentage of CD133(+) (hot spot) in the cancer nest. Ten fields within the hot spot inside the tumor tissue were selected, and CD133 expression was evaluated in 1,000 tumor cells (100 cells per field) at 400× magnification. HIF-1α expression was determined by semi-quantitatively assessing the percentage of stained tumor cells and the staining intensity (SI) (29) (Figure 1C and D). The percentage of positive cells (PP) was rated as follows: 1%-10% PP, +; 11%–50% PP, ++; and >50% PP, +++. SI was scored as weak, +; moderate, ++; and strong, +++. Points for the expression and percentage of PP were calculated using an immunoreactive score (IRS=PP×SI). An IRS score of 0 was considered as negative, a score of 1-2 as low expression, a score of 3-6 as moderate expression, and a score >6 as high expression. Moderate and high expression scores were defined as HIF-1α positive. The evaluation was performed independently by two investigators who were blinded to the clinical findings. Discrepancies between their findings were resolved by discussion.

Figure 1.
Figure 1.

Immunohistochemical detection of CD133 (A, B) and hypoxia-inducible factor 1α (HIF-1α) (C, D). (A) Tumors showing no staining. (B) CD133 expression at the luminal surface and in the cytoplasm. (C) Tumors showing no staining. (D) HIF-1α expression at the luminal surface and in the cytoplasm. Original magnification, ×200 (A-D).

Statistical analyses. The correlations between CD133 and HIF-1α expression and clinicopathological factors were evaluated using unpaired t-tests or chi-squared tests, as appropriate. The unpaired t-test was used for comparing continuous variables, and the chi-squared test was employed for comparison of categorical data. Variables with p-values <0.05 in univariate analyses were subjected to multiple logistic regression analyses. Survival rates and recurrence rates were estimated using the Kaplan–Meier method and were compared using the log-rank test. Variables with p-values <0.05 in univariate analyses were subjected to multivariate Cox proportional hazards analyses. All statistical analyses were performed using the JMP Pro 15.0.0 software (SAS Institute Inc., Cary, NC, USA).

Results

CD133 and HIF-1α expression. The patient characteristics are presented in Table I. Of the 243 enrolled patients, 144 (59.3%) were CD133(+), and 99 (40.7%) were CD133(−). The patients were divided into four groups based on HIF-1α expression: negative, low, moderate, and high, with 84, 36, 93, and 30 participants classified into each group, respectively. The negative and low groups were combined for HIF-1α(−) expression (120 patients), and the moderate and high groups were combined for HIF-1α(+) expression (123 patients).

View this table:
Table I.

Characteristics of rectal cancer patients.

Relationship between CD133 and HIF-1α expression. The positivity rate of CD133 expression was found to increase according to the increase in HIF-1α expression from negative to high, with an increase from 31.0% to 90.0% (p<0.001) (Figure 2). The numbers of patients in each group of combined CD133 and HIF-1α expression were as follows: CD133(−)HIF-1α(−), n=73; CD133(−)HIF-1α(+), n=26; CD133(+)HIF-1α(−), n=47; and CD133(+)HIF-1α(+), n=97.

Figure 2.
Figure 2.

Correlation between HIF-1α expression and CD133 positivity rate. HIF-1α expression significantly correlated with CD133 positivity rate (p<0.001). Tumors were classified into four groups according to HIF-1α expression levels: negative, low, moderate, and high. Negative and low expression levels were defined as HIF-1α negative, and moderate and high expression levels were defined as HIF-1α positive. Black boxes indicate CD133 positive and white boxes indicate CD133 negative.

Relationship between CD133 and HIF-1α expression and clinicopathological features. In the univariate analysis, CD133(+) expression significantly correlated with ypT3-4, positive lymphatic invasion, a pathological tumor regression grade of <2, and lymph node metastasis (Table II). In the multivariate analysis, a pathological tumor regression grade of <2 was the only significant independent variable related to CD133(+) expression (odds ratio, 1.86; p=0.031). Conversely, HIF-1α(+) expression exhibited no association with any clinicopathological factor.

View this table:
Table II.

Correlation between clinicopathological features and CD133 and HIF-1α expression levels.

Correlation between CD133, HIF-1α, and survival. The CD133(+) patients had a significantly shorter relapse-free survival (RFS) than the CD133(−) patients, while no significant difference was observed for overall survival (OS) (CD133(+) vs. CD133(−): 5-year RFS, 63.5% vs. 77.8%; 5-year OS, 81.5% vs. 90.7%, respectively) (Figure 3A and B). Additionally, no significant difference in RFS or OS was observed between HIF-1α(+) and HIF-1α(−) patients [HIF-1α(+) vs. HIF-1α(−): 5-year RFS, 68.6% vs. 70.0%; 5-year OS, 81.8% vs. 88.5%, respectively] (Figure 3C and D).

Figure 3.
Figure 3.

Kaplan–Meier plots showing association of CD133 and HIF-1α expression with prognosis in rectal cancer cases. Relapse-free survival (A, C) and overall survival (B, D) are presented according to CD133 (A, B) and HIF-1α (C, D) expression levels. Blue lines indicate the CD133 or HIF-1α positive groups, and red lines indicate the CD133 or HIF-1α negative groups.

Survival of the four groups based on CD133 and HIF-1α expression. We compared the recurrence rates and prognoses of four groups that were subdivided based on CD133 and HIF-1α expression [CD133(−)HIF-1α(−), CD133(+)HIF-1α(−), CD133(–)HIF-1α(+), and CD133(+)HIF-1α(+)]. The CD133(+)HIF-1α(−) group exhibited poorer RFS and higher liver recurrence rate than the other three groups (Figure 4A and C). No significant difference was observed in OS among the four groups (Figure 4B). Consequently, we divided the patients into two groups: the CD133(+)HIF-1α(−) group and other patients (henceforth referred to as “others”).

Figure 4.
Figure 4.

Kaplan–Meier plots showing the association of combined CD133 and HIF-1α expression with prognosis in rectal cancer cases. Relapse-free survival (A), overall survival (B), liver recurrence rate (C), lung recurrence rate (D), and local recurrence rate (E) are plotted for the combined expression of CD133 and HIF-1α.

Relationship between the CD133(+)HIF-1α(−) group and clinicopathological features. Inclusion in the CD133(+)HIF-1α(−) group significantly correlated with positive vascular invasion and a pathological tumor regression grade of <2 in the univariate analysis (Table III). A pathological tumor regression grade of <2 was the only significant independent variable associated with the CD133(+)HIF-1α(−) group in the multivariate analysis (odds ratio, 2.24; p=0.032).

View this table:
Table III.

Univariate and multivariate analyses of clinicopathological features for the CD133(+)HIF-1α(−) group.

Comparison of survival between the CD133(+)HIF-1α(−) group and the others. The CD133(+)HIF-1α(−) group exhibited significantly shorter RFS than the others, whereas no significant difference in OS was observed [CD133(+)HIF-1α(−) group vs. others: 5-year RFS, 55.1% vs. 73.0%; 5-year OS, 83.3% vs. 85.9%, respectively] (Figure 5A and B). The CD133(+)HIF-1α(−) group exhibited significantly higher liver and local recurrence rates than the others [CD133(+)HIF-1α(−) group vs. others: 5-year liver recurrence rate, 17.3% vs. 5.7%; 5-year local recurrence rate, 15.7% vs. 4.3%, respectively] (Figure 5C and E). There was no significant difference in lung recurrence between the CD133(+)HIF-1α(−) group and the others [CD133(+)HIF-1α(−) group vs. others: 5-year lung recurrence rate, 19.2% vs. 16.1%] (Figure 5D).

Figure 5.
Figure 5.

Kaplan–Meier plot showing association of the CD133(+)HIF-1α(−) group and the others with prognosis. Relapse-free survival (A), overall survival (B), liver recurrence rate (C), lung recurrence rate (D), and local recurrence rate (E) are plotted for the CD133(+)HIF-1α(−) group and the others. Blue lines indicate the CD133(+)HIF-1α(−) group, and red lines indicate the others.

Multivariate analyses between RFS, liver recurrence rate, and local recurrence rate and clinicopathological features. The CD133(+)HIF-1α(−) group had poor RFS in the multivariate analysis [hazard ratio (HR)=1.63; p=0.071] (Table IV). For liver recurrence, CD133(+)HIF-1α(−) expression was recognized as an independent risk factor in the multivariate analysis (HR, 2.97; p=0.022). Additionally, CD133(+)HIF-1α(−) expression correlated with higher local recurrence in the multivariate analysis (HR=2.74; p=0.055).

View this table:
Table IV.

Univariate and multivariate analyses of prognostic valuables for relapse-free survival, liver recurrence, and local recurrence after curative resection.

Discussion

In this study, we evaluated the protein expression of CD133 and HIF-1α by immunohistochemistry for patients with advanced low rectal cancer who received CRT followed by curative resection. We investigated interactions between CD133 and HIF-1α and the relationship between these proteins and prognosis. The CD133 positivity rate after CRT increased according to the increase in HIF-1α expression (Figure 2). Additionally, most CD133(−) patients were HIF-1α(−), and most CD133(+) patients were HIF-1α(+). These two groups collectively accounted for 70% of all patients (Table I). Although it has previously been reported that high HIF-1α expression coincided with high CD133 expression in renal cell carcinoma (30) and intrahepatic cholangiocarcinoma (31) cases, ours was the first study of colorectal cancer. CD133 and HIF-1α seemed to be co-regulated in previous studies given that CD133 knockdown depressed HIF-1α expression (20) and HIF-1α knockdown depressed CD133 expression (21). Our results seem to corroborate these interactions.

With regard to the relationship between CD133 expression and clinicopathological factors or prognosis, the CD133(+) group had a poor tumor regression grade that was statistically significant (Table II), but there was no correlation with prognosis (Figure 3) consistent with our previous findings (8, 11). Our study showed no correlation between HIF-1α expression and clinicopathological factors or prognosis (Table II, Figure 3), which differs from previous reports (17–19). Altogether, the prognostic impacts of either CD133 or HIF-1α expression alone were not strong. However, the combinatorial assessment of these proteins revealed that the CD133(+)HIF-1α(−) group had a poorer tumor regression grade, three times more liver recurrence and local recurrence, and poorer prognosis compared to the other patients (Table III and Table IV, Figure 5).

Previous cell culture experiments (32, 33) and a study using clinical specimens (10) showed that CD133(+) cells had higher invasive activity, higher metastaticity, and poorer prognosis than CD133(−) cells in different types of cancer. Additionally, it was reported that CD133 expression was increased by radiation (5–8). By contrast, it was also reported that radiation increased HIF-1α expression in cell culture experiments (14–16) whereas radiation decreased HIF-1α expression in resected specimens (17, 19, 34). This discrepancy could be explained by the fact that HIF-1α expression increased early after radiation, while tumor regression and reoxygenation resulted in the degradation of HIF-1α over the next few days (17). HIF-1α expression could have decreased in resected specimens during the waiting time for surgery, which was one month or more from the completion of radiotherapy. For the above reasons, resected specimens tend to become CD133(+)HIF-1α(−) after CRT, and continuous up-regulation of HIF-1α by CD133 should be necessary to have CD133(+)HIF-1α(+) expression.

The characteristics of HIF-1α expression, which suppresses tumor proliferation, were first observed in renal cancer cells (35) and have been subsequently reported in many cancer cells such as acute myeloid leukemia (36) and glioma cells (37). Tiwari et al. showed that HIF-1α played a role as a tumor suppressor by preventing degradation of p53 protein in pancreatic cancer cell lines, and the loss of HIF-1α increased the invasion and metastasis of pancreatic cancer (38). These reports indicated that the loss of HIF-1α expression increases metastasis and recurrence. Although the CD133(+) group had higher invasive activity and metastaticity than the CD133(−) group, HIF-1α expression in this CD133(+) group should have a better prognosis because HIF-1α expression suppresses tumor invasion and metastasis. By contrast, the CD133(+) without HIF-1α expression was considered to be the consequence of low activation of HIF-1α by CD133, resulting in the lack of metastasis-suppression by HIF-1α and a markedly higher rate of distant metastasis and cancer recurrence.

Although patients in the CD133(+)HIF-1α(−) group had more lung recurrence than the others until one year after resection, the difference was marginal at five years post-resection (Figure 5D). However, data were insufficient because of the small number of patients.

Our study has several limitations. Because this is a retrospective study from a single institution, the number of patients enrolled was relatively small. In addition, precise assessment of how CD133 and HIF-1α expression changed after CRT and how these proteins affected tumor recurrence still remain to be investigated. Further investigation on molecular mechanisms is warranted.

In conclusion, CD133(+)HIF-1α(−) expression after CRT is a promising marker for recurrence, particularly for liver recurrence. Patients with CD133(+)HIF-1α(−) expression had a poor prognosis, and therefore intensive surveillance and strong adjuvant chemotherapy should be performed for these patients.

Acknowledgements

This study was supported by Grants-in-Aid for Scientific Research (C: grant number; 18K07194, C: grant number; 19K09114, C: grant number; 19K09115, C: grant number; 20K09051, Challenging Research (Exploratory): grant number; 20K21626, B: grant number; 21H02778) from the Japan Society for the promotion of Science (Tokyo, Japan). This research is supported by the Project for Cancer Research and Therapeutic Evolution (P-CREATE), grant number: JP 19cm0106502 from the Japan Agency for Medical Research and Development (AMED) (Tokyo, Japan).

Footnotes

  • Authors’ Contributions

    SI and KK conceptualized the study. YT and KK designed and coordinated the study and wrote the manuscript. KO, JK, and YY collected and analyzed data. SA, YN, HA, and HS interpreted data. HN, KS, KM, SE, and SI provided critical revisions. All Authors have read and approved the final version of the manuscript.

  • Conflicts of Interest

    The Authors declare no conflicts of interest in relation to this study.

  • Received January 14, 2022.
  • Revision received February 20, 2022.
  • Accepted February 21, 2022.

References

  1. ↵
    1. Benson AB 3rd.,
    2. Venook AP,
    3. Bekaii-Saab T,
    4. Chan E,
    5. Chen YJ,
    6. Cooper HS,
    7. Engstrom PF,
    8. Enzinger PC,
    9. Fenton MJ,
    10. Fuchs CS,
    11. Grem JL,
    12. Grothey A,
    13. Hochster HS,
    14. Hunt S,
    15. Kamel A,
    16. Kirilcuk N,
    17. Leong LA,
    18. Lin E,
    19. Messersmith WA,
    20. Mulcahy MF,
    21. Murphy JD,
    22. Nurkin S,
    23. Rohren E,
    24. Ryan DP,
    25. Saltz L,
    26. Sharma S,
    27. Shibata D,
    28. Skibber JM,
    29. Sofocleous CT,
    30. Stoffel EM,
    31. Stotsky-Himelfarb E,
    32. Willett CG,
    33. Gregory KM and
    34. Freedman-Cass D
    : Rectal cancer, version 2.2015. J Natl Compr Canc Netw 13(6): 719-28; quiz 728, 2015. PMID: 26085388. DOI: 10.6004/jnccn.2015.0087
    OpenUrlAbstract/FREE Full Text
  2. ↵
    1. Glynne-Jones R,
    2. Wyrwicz L,
    3. Tiret E,
    4. Brown G,
    5. Rödel C,
    6. Cervantes A,
    7. Arnold D and ESMO Guidelines Committee
    : Rectal cancer: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Ann Oncol 28(suppl_4): iv22-iv40, 2017. PMID: 28881920. DOI: 10.1093/annonc/mdx224
    OpenUrlCrossRefPubMed
  3. ↵
    1. O’Brien CA,
    2. Pollett A,
    3. Gallinger S and
    4. Dick JE
    : A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. Nature 445(7123): 106-110, 2007. PMID: 17122772. DOI: 10.1038/nature05372
    OpenUrlCrossRefPubMed
  4. ↵
    1. Ricci-Vitiani L,
    2. Lombardi DG,
    3. Pilozzi E,
    4. Biffoni M,
    5. Todaro M,
    6. Peschle C and
    7. De Maria R
    : Identification and expansion of human colon-cancer-initiating cells. Nature 445(7123): 111-115, 2007. PMID: 17122771. DOI: 10.1038/nature05384
    OpenUrlCrossRefPubMed
  5. ↵
    1. Kawamoto A,
    2. Tanaka K,
    3. Saigusa S,
    4. Toiyama Y,
    5. Morimoto Y,
    6. Fujikawa H,
    7. Iwata T,
    8. Matsushita K,
    9. Yokoe T,
    10. Yasuda H,
    11. Inoue Y,
    12. Miki C and
    13. Kusunoki M
    : Clinical significance of radiation-induced CD133 expression in residual rectal cancer cells after chemoradiotherapy. Exp Ther Med 3(3): 403-409, 2012. PMID: 22969903. DOI: 10.3892/etm.2011.438
    OpenUrlCrossRefPubMed
  6. ↵
    1. Saigusa S,
    2. Tanaka K,
    3. Toiyama Y,
    4. Yokoe T,
    5. Okugawa Y,
    6. Ioue Y,
    7. Miki C and
    8. Kusunoki M
    : Correlation of CD133, OCT4, and SOX2 in rectal cancer and their association with distant recurrence after chemoradiotherapy. Ann Surg Oncol 16(12): 3488-3498, 2009. PMID: 19657699. DOI: 10.1245/s10434-009-0617-z
    OpenUrlCrossRefPubMed
  7. ↵
    1. Saigusa S,
    2. Tanaka K,
    3. Toiyama Y,
    4. Yokoe T,
    5. Okugawa Y,
    6. Kawamoto A,
    7. Yasuda H,
    8. Morimoto Y,
    9. Fujikawa H,
    10. Inoue Y,
    11. Miki C and
    12. Kusunoki M
    : Immunohistochemical features of CD133 expression: association with resistance to chemoradiotherapy in rectal cancer. Oncol Rep 24(2): 345-350, 2010. PMID: 20596619. DOI: 10.3892/or_00000865
    OpenUrlCrossRefPubMed
  8. ↵
    1. Hongo K,
    2. Kazama S,
    3. Sunami E,
    4. Tsuno NH,
    5. Takahashi K,
    6. Nagawa H and
    7. Kitayama J
    : Immunohistochemical detection of CD133 is associated with tumor regression grade after chemoradiotherapy in rectal cancer. Med Oncol 29(4): 2849-2857, 2012. PMID: 22246526. DOI: 10.1007/s12032-012-0161-8
    OpenUrlCrossRefPubMed
    1. Hiroishi K,
    2. Inomata M,
    3. Kashima K,
    4. Yasuda K,
    5. Shiraishi N,
    6. Yokoyama S and
    7. Kitano S
    : Cancer stem cell-related factors are associated with the efficacy of pre-operative chemoradiotherapy for locally advanced rectal cancer. Exp Ther Med 2(3): 465-470, 2011. PMID: 22977526. DOI: 10.3892/etm.2011.243
    OpenUrlCrossRefPubMed
  10. ↵
    1. Sprenger T,
    2. Conradi LC,
    3. Beissbarth T,
    4. Ermert H,
    5. Homayounfar K,
    6. Middel P,
    7. Rüschoff J,
    8. Wolff HA,
    9. Schüler P,
    10. Ghadimi BM,
    11. Rödel C,
    12. Becker H,
    13. Rödel F and
    14. Liersch T
    : Enrichment of CD133-expressing cells in rectal cancers treated with preoperative radiochemotherapy is an independent marker for metastasis and survival. Cancer 119(1): 26-35, 2013. PMID: 22736392. DOI: 10.1002/cncr.27703
    OpenUrlCrossRefPubMed
  11. ↵
    1. Harada Y,
    2. Kazama S,
    3. Morikawa T,
    4. Murono K,
    5. Yasuda K,
    6. Otani K,
    7. Nishikawa T,
    8. Tanaka T,
    9. Kiyomatsu T,
    10. Kawai K,
    11. Hata K,
    12. Nozawa H,
    13. Yamaguchi H,
    14. Ishihara S and
    15. Watanabe T
    : Leucine-rich repeat-containing G protein-coupled receptor 5 and CD133 expression is associated with tumor progression and resistance to preoperative chemoradiotherapy in low rectal cancer. Oncol Lett 14(6): 7791-7798, 2017. PMID: 29250176. DOI: 10.3892/ol.2017.7207
    OpenUrlCrossRefPubMed
  12. ↵
    1. Vaupel P,
    2. Höckel M and
    3. Mayer A
    : Detection and characterization of tumor hypoxia using pO2 histography. Antioxid Redox Signal 9(8): 1221-1235, 2007. PMID: 17536958. DOI: 10.1089/ars.2007.1628
    OpenUrlCrossRefPubMed
  13. ↵
    1. Semenza GL
    : Targeting HIF-1 for cancer therapy. Nat Rev Cancer 3(10): 721-732, 2003. PMID: 13130303. DOI: 10.1038/nrc1187
    OpenUrlCrossRefPubMed
  14. ↵
    1. Miyasaka A,
    2. Oda K,
    3. Ikeda Y,
    4. Sone K,
    5. Fukuda T,
    6. Inaba K,
    7. Makii C,
    8. Enomoto A,
    9. Hosoya N,
    10. Tanikawa M,
    11. Uehara Y,
    12. Arimoto T,
    13. Kuramoto H,
    14. Wada-Hiraike O,
    15. Miyagawa K,
    16. Yano T,
    17. Kawana K,
    18. Osuga Y and
    19. Fujii T
    : PI3K/mTOR pathway inhibition overcomes radioresistance via suppression of the HIF1-α/VEGF pathway in endometrial cancer. Gynecol Oncol 138(1): 174-180, 2015. PMID: 25913131. DOI: 10.1016/j.ygyno.2015.04.015
    OpenUrlCrossRefPubMed
    1. Lu H,
    2. Liang K,
    3. Lu Y and
    4. Fan Z
    : The anti-EGFR antibody cetuximab sensitizes human head and neck squamous cell carcinoma cells to radiation in part through inhibiting radiation-induced upregulation of HIF-1α. Cancer Lett 322(1): 78-85, 2012. PMID: 22348829. DOI: 10.1016/j.canlet.2012.02.012
    OpenUrlCrossRefPubMed
  16. ↵
    1. Okuno T,
    2. Kawai K,
    3. Hata K,
    4. Murono K,
    5. Emoto S,
    6. Kaneko M,
    7. Sasaki K,
    8. Nishikawa T,
    9. Tanaka T and
    10. Nozawa H
    : SN-38 acts as a radiosensitizer for colorectal cancer by inhibiting the radiation-induced up-regulation of HIF-1α. Anticancer Res 38(6): 3323-3331, 2018. PMID: 29848680. DOI: 10.21873/anticanres.12598
    OpenUrlAbstract/FREE Full Text
  17. ↵
    1. Havelund BM,
    2. Sørensen FB,
    3. Pløen J,
    4. Lindebjerg J,
    5. Spindler KL and
    6. Jakobsen A
    : Immunohistological expression of HIF-1α, GLUT-1, Bcl-2 and Ki-67 in consecutive biopsies during chemoradiotherapy in patients with rectal cancer. APMIS 121(2): 127-138, 2013. PMID: 23030583. DOI: 10.1111/j.1600-0463.2012.02949.x
    OpenUrlCrossRefPubMed
  18. ↵
    1. Saigusa S,
    2. Tanaka K,
    3. Toiyama Y,
    4. Yokoe T,
    5. Okugawa Y,
    6. Koike Y,
    7. Fujikawa H,
    8. Inoue Y,
    9. Miki C and
    10. Kusunoki M
    : Clinical significance of CD133 and hypoxia inducible factor-1α gene expression in rectal cancer after preoperative chemoradiotherapy. Clin Oncol (R Coll Radiol) 23(5): 323-332, 2011. PMID: 20970309. DOI: 10.1016/j.clon.2010.09.012
    OpenUrlCrossRefPubMed
  19. ↵
    1. Korkeila E,
    2. Jaakkola PM,
    3. Syrjänen K,
    4. Sundström J and
    5. Pyrhönen S
    : Preoperative radiotherapy downregulates the nuclear expression of hypoxia-inducible factor-1alpha in rectal cancer. Scand J Gastroenterol 45(3): 340-348, 2010. PMID: 20001759. DOI: 10.3109/00365520903483635
    OpenUrlCrossRefPubMed
  20. ↵
    1. Maeda K,
    2. Ding Q,
    3. Yoshimitsu M,
    4. Kuwahata T,
    5. Miyazaki Y,
    6. Tsukasa K,
    7. Hayashi T,
    8. Shinchi H,
    9. Natsugoe S and
    10. Takao S
    : CD133 modulate HIF-1α expression under hypoxia in EMT phenotype pancreatic cancer stem-like cells. Int J Mol Sci 17(7): 1025, 2016. PMID: 27367674. DOI: 10.3390/ijms17071025
    OpenUrlCrossRefPubMed
  21. ↵
    1. Soeda A,
    2. Park M,
    3. Lee D,
    4. Mintz A,
    5. Androutsellis-Theotokis A,
    6. McKay RD,
    7. Engh J,
    8. Iwama T,
    9. Kunisada T,
    10. Kassam AB,
    11. Pollack IF and
    12. Park DM
    : Hypoxia promotes expansion of the CD133-positive glioma stem cells through activation of HIF-1alpha. Oncogene 28(45): 3949-3959, 2009. PMID: 19718046. DOI: 10.1038/onc.2009.252
    OpenUrlCrossRefPubMed
  22. ↵
    1. Okada M,
    2. Kawai K,
    3. Sonoda H,
    4. Shiratori H,
    5. Kishikawa J,
    6. Nagata H,
    7. Nozawa H,
    8. Sasaki K,
    9. Kaneko M,
    10. Murono K,
    11. Emoto S,
    12. Iida Y,
    13. Ishii H,
    14. Yokoyama Y,
    15. Anzai H,
    16. Hasegawa K and
    17. Ishihara S
    : Epithelial-mesenchymal transition and metastatic ability of CD133+ colorectal cancer stem-like cells under hypoxia. Oncol Lett 21(1): 19, 2021. PMID: 33240425. DOI: 10.3892/ol.2020.12280
    OpenUrlCrossRefPubMed
  23. ↵
    1. Brierley JD,
    2. Gospodarowicz MK and
    3. Wittekind C
    : TNM Classification of malignant tumours. 8th ed. New Jersey, John Wiley & Sons, Inc, 2017.
  24. ↵
    1. Fritz A,
    2. Percy C,
    3. Jack A,
    4. Shanmugaratnam K,
    5. Sobin L,
    6. Parkin DM and
    7. Whelan S
    : International Classification of Diseases for Oncology. 3rd ed. Geneva, World Health Organization, 2000. Available at: https://apps.who.int/iris/handle/10665/42344 [Last accessed on January 10, 2022]
  25. ↵
    1. Japanese Society for Cancer of the Colon and Rectum
    : Japanese Classification of colorectal, appendiceal, and anal carcinoma: the 3d English edition [Secondary Publication]. J Anus Rectum Colon 3(4): 175-195, 2019. PMID: 31768468. DOI: 10.23922/jarc.2019-018
    OpenUrlCrossRefPubMed
  26. ↵
    1. Maeda S,
    2. Shinchi H,
    3. Kurahara H,
    4. Mataki Y,
    5. Maemura K,
    6. Sato M,
    7. Natsugoe S,
    8. Aikou T and
    9. Takao S
    : CD133 expression is correlated with lymph node metastasis and vascular endothelial growth factor-C expression in pancreatic cancer. Br J Cancer 98(8): 1389-1397, 2008. PMID: 18349830. DOI: 10.1038/sj.bjc.6604307
    OpenUrlCrossRefPubMed
    1. Li CY,
    2. Li BX,
    3. Liang Y,
    4. Peng RQ,
    5. Ding Y,
    6. Xu DZ,
    7. Zhang X,
    8. Pan ZZ,
    9. Wan DS,
    10. Zeng YX,
    11. Zhu XF and
    12. Zhang XS
    : Higher percentage of CD133+ cells is associated with poor prognosis in colon carcinoma patients with stage IIIB. J Transl Med 7: 56, 2009. PMID: 19583834. DOI: 10.1186/1479-5876-7-56
    OpenUrlCrossRefPubMed
  28. ↵
    1. Lugli A,
    2. Iezzi G,
    3. Hostettler I,
    4. Muraro MG,
    5. Mele V,
    6. Tornillo L,
    7. Carafa V,
    8. Spagnoli G,
    9. Terracciano L and
    10. Zlobec I
    : Prognostic impact of the expression of putative cancer stem cell markers CD133, CD166, CD44s, EpCAM, and ALDH1 in colorectal cancer. Br J Cancer 103(3): 382-390, 2010. PMID: 20606680. DOI: 10.1038/sj.bjc.6605762
    OpenUrlCrossRefPubMed
  29. ↵
    1. Fan LF,
    2. Dong WG,
    3. Jiang CQ,
    4. Qian Q and
    5. Yu QF
    : Role of Hypoxia-inducible factor-1 alpha and Survivin in colorectal carcinoma progression. Int J Colorectal Dis 23(11): 1057-1064, 2008. PMID: 18704463. DOI: 10.1007/s00384-008-0511-3
    OpenUrlCrossRefPubMed
  30. ↵
    1. Sun C,
    2. Song H,
    3. Zhang H,
    4. Hou C,
    5. Zhai T,
    6. Huang L and
    7. Zhang L
    : CD133 expression in renal cell carcinoma (RCC) is correlated with nuclear hypoxia-inducing factor 1α (HIF-1α). J Cancer Res Clin Oncol 138(10): 1619-1624, 2012. PMID: 22614155. DOI: 10.1007/s00432-012-1237-8
    OpenUrlCrossRefPubMed
  31. ↵
    1. Shimada M,
    2. Sugimoto K,
    3. Iwahashi S,
    4. Utsunomiya T,
    5. Morine Y,
    6. Imura S and
    7. Ikemoto T
    : CD133 expression is a potential prognostic indicator in intrahepatic cholangiocarcinoma. J Gastroenterol 45(8): 896-902, 2010. PMID: 20379837. DOI: 10.1007/s00535-010-0235-3
    OpenUrlCrossRefPubMed
  32. ↵
    1. Ding Q,
    2. Yoshimitsu M,
    3. Kuwahata T,
    4. Maeda K,
    5. Hayashi T,
    6. Obara T,
    7. Miyazaki Y,
    8. Matsubara S,
    9. Natsugoe S and
    10. Takao S
    : Establishment of a highly migratory subclone reveals that CD133 contributes to migration and invasion through epithelial-mesenchymal transition in pancreatic cancer. Hum Cell 25(1): 1-8, 2012. PMID: 22109279. DOI: 10.1007/s13577-011-0037-9
    OpenUrlCrossRefPubMed
  33. ↵
    1. Zhang H,
    2. Yang N,
    3. Sun B,
    4. Jiang Y,
    5. Hou C,
    6. Ji C,
    7. Zhang Y,
    8. Liu Y and
    9. Zuo P
    : CD133 positive cells isolated from A549 cell line exhibited high liver metastatic potential. Neoplasma 61(2): 153-160, 2014. PMID: 24299311. DOI: 10.4149/neo_2014_021
    OpenUrlCrossRefPubMed
  34. ↵
    1. Kim HJ,
    2. Bae SB,
    3. Jeong D,
    4. Kim ES,
    5. Kim CN,
    6. Park DG,
    7. Ahn TS,
    8. Cho SW,
    9. Shin EJ,
    10. Lee MS and
    11. Baek MJ
    : Upregulation of stromal cell-derived factor 1α expression is associated with the resistance to neoadjuvant chemoradiotherapy of locally advanced rectal cancer: angiogenic markers of neoadjuvant chemoradiation. Oncol Rep 32(6): 2493-2500, 2014. PMID: 25241658. DOI: 10.3892/or.2014.3504
    OpenUrlCrossRefPubMed
  35. ↵
    1. Shen C,
    2. Beroukhim R,
    3. Schumacher SE,
    4. Zhou J,
    5. Chang M,
    6. Signoretti S and
    7. Kaelin WG Jr.
    : Genetic and functional studies implicate HIF1α as a 14q kidney cancer suppressor gene. Cancer Discov 1(3): 222-235, 2011. PMID: 22037472. DOI: 10.1158/2159-8290.CD-11-0098
    OpenUrlCrossRefPubMed
  36. ↵
    1. Velasco-Hernandez T,
    2. Hyrenius-Wittsten A,
    3. Rehn M,
    4. Bryder D and
    5. Cammenga J
    : HIF-1α can act as a tumor suppressor gene in murine acute myeloid leukemia. Blood 124(24): 3597-3607, 2014. PMID: 25267197. DOI: 10.1182/blood-2014-04-567065
    OpenUrlAbstract/FREE Full Text
  37. ↵
    1. Koivunen P,
    2. Lee S,
    3. Duncan CG,
    4. Lopez G,
    5. Lu G,
    6. Ramkissoon S,
    7. Losman JA,
    8. Joensuu P,
    9. Bergmann U,
    10. Gross S,
    11. Travins J,
    12. Weiss S,
    13. Looper R,
    14. Ligon KL,
    15. Verhaak RG,
    16. Yan H and
    17. Kaelin WG Jr.
    : Transformation by the (R)-enantiomer of 2-hydroxyglutarate linked to EGLN activation. Nature 483(7390): 484-488, 2012. PMID: 22343896. DOI: 10.1038/nature10898
    OpenUrlCrossRefPubMed
  38. ↵
    1. Tiwari A,
    2. Tashiro K,
    3. Dixit A,
    4. Soni A,
    5. Vogel K,
    6. Hall B,
    7. Shafqat I,
    8. Slaughter J,
    9. Param N,
    10. Le A,
    11. Saunders E,
    12. Paithane U,
    13. Garcia G,
    14. Campos AR,
    15. Zettervall J,
    16. Carlson M,
    17. Starr TK,
    18. Marahrens Y,
    19. Deshpande AJ,
    20. Commisso C,
    21. Provenzano PP and
    22. Bagchi A
    : Loss of HIF1A from pancreatic cancer cells increases expression of PPP1R1B and degradation of p53 to promote invasion and metastasis. Gastroenterology 159(5): 1882-1897.e5, 2020. PMID: 32768595. DOI: 10.1053/j.gastro.2020.07.046
    OpenUrlCrossRefPubMed
Back to top

In this issue

Print
Download PDF
Article Alerts
Email Article
Citation Tools
Reprints and Permissions
CD133(+)HIF-1α(−) Expression After Chemoradiotherapy Predicts Poor Prognosis in Rectal Cancer
YUICHI TACHIKAWA, KAZUSHIGE KAWAI, KOSUKE OZAKI, HIROAKI NOZAWA, KAZUHITO SASAKI, KOJI MURONO, SHIGENOBU EMOTO, JUNKO KISHIKAWA, YUICHIRO YOKOYAMA, SHINYA ABE, YUZO NAGAI, HIROYUKI ANZAI, HIROFUMI SONODA, SOICHIRO ISHIHARA
Anticancer Research Apr 2022, 42 (4) 2033-2043; DOI: 10.21873/anticanres.15684
Twitter logo Facebook logo Mendeley logo

Jump to section

Keywords