Research ArticleExperimental Studies
Open Access

PAR1 Is a Candidate Target for the Treatment of Peritoneal Dissemination in Gastric Cancer

DAISUKE FUJIMOTO and HIROTOSHI KOBAYASHI
Anticancer Research November 2024, 44 (11) 4857-4867; DOI: https://doi.org/10.21873/anticanres.17311

Abstract

Background/Aim: Peritoneal dissemination (PD) is a frequent cause of death in gastric cancer (GC), and there is evidence of an association between protease-activated receptor-1 (PAR1) and the development of PD. This study hypothesized that PD in GC might be influenced by PAR1. Materials and Methods: The cytotoxic effect of paclitaxel (PTX) on PAR1-transfected MKN45 (MKN45/PAR1) cells was analyzed using the MTT assay, and IC50 values were determined. In female athymic nude mice, MKN45/PAR1 cells were suspended in 0.05 ml phosphate-buffered saline (PBS) medium and inoculated into the stomach mid-wall. In each group, intraperitoneal injections of PBS, PTX, SCH79797 (PAR1-antagonist), or PTX plus SCH79797 were administered on days 8, 15, and 22 following tumor inoculation. At 56 days after tumor inoculation, mice were examined for both abdominal tumor nodule status and size and weight of the tumors. Results: The IC50 of PTX for MKN45/PAR1 cells was 0.0697 μM and that of SCH79797 was 0.0145 μM. Mean survival of the MKN45/PAR1 mice in the PBS group was 28.75 days, whereas survival times for the mice treated with SCH79797, PTX, or a combination of PTX and SCH79797 were 31.2, 49.2, and 48.5 days, respectively. Tumor weight was smaller in the group receiving PTX and SCH79797 intraperitoneally compared with that in the PBS group (1,086±127.2 mg vs. 33.2±19.9 mg; p<0.001). Conclusion: The PAR1 antagonist was found to inhibit PD in a PAR1-expressing GC cell line. PAR1 may serve as a promising therapeutic target for managing PD in gastric cancer, as it plays a crucial role in its progression.

Key Words:

Despite improvements in surgical treatment and chemotherapy, the prognosis of gastric cancer at an advanced stage is still poor. Practically, peritoneal carcinomatosis is a major cause of death in advanced gastric cancer and often occurs after surgery (1, 2). Currently, there is no effective therapy for this condition. The 5-year survival rate of patients with peritoneal carcinomatosis is only 2% even when including patients with intraperitoneal free cancer cells without macroscopic peritoneal carcinomatosis (3).

Protease-activated receptor 1 (PAR1) has been found to be instrumentally involved in cell growth and invasion of tumor-derived cells (4, 5). In addition to regulating cell function via the PARs, thrombin may also affect cell function via the activation of metalloproteinase-2 (MMP2) (6, 7). Apart from serine proteinases that can activate PARs to affect cancer cell behavior, MMPs have been known to be involved in cancer metastasis and invasion (8-12). Surprisingly, similar to thrombin, MMP1 has been observed to regulate invasion and tumorigenesis of breast cancer-derived cells by a process involving PAR1 (13), thus providing an important link between tumor-generated metalloproteinases and PAR signaling. In addition to PAR overexpression, breast carcinoma cells display aberrant PAR1 trafficking, which causes persistent signaling and cellular invasion (14). PAR1 over-expression has also been documented in prostate cancer-derived cells and has been linked to activation of NF-Embedded ImageB, with an increase in NF-Embedded ImageB-regulated inflammatory cytokines, such as IL-6 and IL-8 (5, 15).

In our previous work, we showed that activated PAR1 triggered proliferation and invasion in gastric cancer cells via NF-Embedded ImageB, tenascin-C, and epidermal growth factor receptor, and using an immunohistochemical approach with gastric carcinoma tissue, we found that the expression of PAR1 was associated with peritoneal dissemination and poorer prognosis compared with expression-negative tumors (16-20). Recent studies of gastric cancer and PAR1 have also reported that PAR1 is continuously involved in gastritis associated with Helicobacter pylori infection which is implicated in gastric carcinogenesis (21, 22). Furthermore, the long non-coding RNA ncRuPAR, which is a non-protein-coding RNA upstream of coagulation factor II thrombin receptor/PAR1, regulates the proliferation of gastric cancer cells and promotes their apoptosis potentially by inhibiting PAR1 (23).

We hypothesized that PAR1 might be involved in the development of peritoneal carcinomatosis caused by gastric cancer. Here, we report evidence that PAR1 plays a role in the development of peritoneal carcinomatosis from gastric cancer and could be an attractive target as a novel therapeutic approach against peritoneal carcinomatosis of gastric cancer.

Materials and Methods

Chemicals, drugs, and supplies. Roswell Park Memorial institute (RPMI) 1640 medium and fetal bovine serum were purchased from GIBCO BRL (Tokyo, Japan), and BALB/cAJcl-nu nude mice were from Nippon Korea (Tokyo, Japan). Paclitaxel (PTX) was obtained from Sigma-Aldrich (St. Louis, MO, USA), and the selective PAR1 antagonist SCH79797 [50% inhibitory concentration (IC50)=70 nM] was purchased from Tocris Bioscience (Avon Mouth, UK). PTX and SCH79797 were dissolved with ethanol.

Cell lines. MKN45 human gastric cancer cells were purchased from the Riken Cell Bank (Tsukuba, Japan). MKN45/PAR1 cells transfected with a PAR1-incorporated plasmid and MKN45/mock cells transfected with an empty plasmid, which were created in a previous experiment, were also used (17). Cells were cultured in RPMI 1640 medium supplemented with 10% heat-inactivated fetal bovine serum plus 1% penicillin/streptomycin and 1% glutamine. Cultures were maintained in a 5% CO2 humidified incubator at 37°C, and experiments were performed with exponentially growing cells.

Cytotoxicity assay. The 3-(4,5-dimethiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was performed as previously described (24). Cell mixtures were adjusted to 5×104 cells/ml and seeded at 50 μl/well in a 96-well microplate reader (BD FALCON, Schaffhausen, Switzerland). After incubation in a 5% CO2 incubator at 37°C for 24 h, 50 μl/well of the drug solution adjusted to the final concentration was added, and the drug was incubated with the cultured cells in the CO2 incubator for 72 h. In this study, we measured the IC50 of PTX against MKN45/PAR1 cells and the IC50 of PTX under three conditions: with 30 nM, 70 nM, and 150 nM of SCH79797 added to the culture medium. Before the end of the reaction, 10 μl/well of 0.5% MTT-PBS (phosphate-buffered saline) solution was added to 100 μl of culture medium to make it luminescent for 4 h. The culture medium was then removed, the cells were lysed by adding 200 μl/well of dimethyl sulfoxide, and absorbance at 590 nm was measured in a microplate reader (Thermo Fisher Scientific, Waltham, MA, USA). Cell viability was expressed as the ratio of absorbance of the drug-treated group to that of the drug-untreated group. The IC50 was calculated using Origin 8.1J (OriginLab, Northampton, MA, USA).

Animal studies. Pathogen-free 6-week-old female BALB/cAJcl-nu nude mice were housed under specific pathogen-free conditions and given sterile food and autoclaved water. The animals were maintained in an animal facility in a 12-hour light-dark cycle at a temperature (20°C) and humidity (50%) controlled environment. Food and water were freely available. Body weights were also measured twice per week. All mice had a mean body weight of 21±0.4 g at the start of treatment, complying with the standards set in the Guidelines for the Care and Use of Laboratory Animals in the University of Fukui. The mice were anesthetized by intraperitoneal injection of isoflurane (Sigma-Aldrich) at a dose of 0.28 mg/g body weight for implantation of cells into the stomach wall of each mouse. Next, after a small incision was made in the midline of the abdomen of the anesthetized mice, 2×106 MKN45/PAR1 or MKN45/mock cells suspended in a 0.05-ml volume of PBS medium were inoculated into the mid-wall of the greater curvature in the gastric gland area in each of 25 mice using a 27-gauge needle (Nipro, Tokyo, Japan). The stomach was then returned into the peritoneal cavity, and the abdominal wall and skin were closed with 3-0 VICRYL (ETHICON, Inc., Somerville, NJ, USA) (25). For the 25 mice inoculated with MKN45/PAR1 or MKN45/mock cells into the stomach, starting one week after administration, the first group of 5 mice received 0.04 μM PTX once weekly on days 8, 15, and 22; the second group of 5 mice received 70 nM SCH79797 once weekly on days 8, 15, and 22; the third group of 5 mice received 0.04 μM PTX and 70 nM SCH79797 simultaneously once weekly on days 8, 15, and 22; the fourth group of 5 mice received 500 μl PBS once weekly on days 8, 15, and 22; and the fifth group of 5 mice were kept for eight weeks without any treatment. The control groups for mouse weight comparison in the MKN45/PAR1 group and MKN45/mock group each comprised 5 mice, 1 mouse for each different drug dose, which were simply fed food without injection of cancer cells. The body weights of the mice were routinely measured to monitor the extent of the development of peritoneal carcinomatosis. Humane endpoints were reached when tumors metastasized or led to rapid body weight loss (>20%), or signs of immobility, a huddled posture, inability to eat, ruffled fur, self-mutilation, ulceration, infection or necrosis were observed. Mice still alive at 56 days after tumor inoculation into the stomach were intraperitoneally administered 0.15 mg pentobarbital (Sigma-Aldrich) per mg mouse weight after death was confirmed sacrificed and necropsied after death confirmation. Death was confirmed by monitoring for 10 min after cardiac arrest was recognized by at least two investigators. The extent of peritoneal carcinomatosis was then quantified using the Peritoneal Cancer Index (PCI) (26) adapted to tumor sizes in mice (27). The weight of the total peritoneal tumor burden was measured for each individual mouse.

Statistical analysis. Results are presented as the mean±SE. Differences in continuous variables between the two groups were evaluated using the Student’s t-test and the Mann–Whitney U-test for non-parametric values. Differences in continuous variables between three or more groups were evaluated using analysis of variance, and the Dunnett test for parametric values or Kruskal–Wallis test and Dunn–Bonferroni test for non-parametric values was performed. Overall survival curves were plotted according to the Kaplan–Meier method and the log-rank test was used to compare the survival curves. Differences between groups were considered statistically significant at p<0.05. Statistical analyses were carried out using the computer software package SPSS Statistics v28 (IBM, Chicago, IL, USA). Experiments were performed at least three times.

Results

Inhibitory effect of PTX on tumor growth of MKN45/PAR1 cells. To assess the inhibitory effect of PTX on MKN45/PAR1 cells in vitro, the MTT assay was conducted. Based on the result of the preliminary experiment (Figure 1A), the exposure time of MKN45/PAR1 cells to PTX was set at 72 h, and after 72 h of exposure, these cells were washed and fixed to measure cell growth indirectly. From the preliminary experimental result (Figure 1A), on the third day of the experiment (i.e., at 72 h of exposure to PTX), cells were washed and fixed, and cell growth was determined indirectly. As shown in Figure 2, the PTX IC50 value for MKN45/PAR1 cells in the presence PTX for 72 h was 0.0697±0.0125 μM.

Figure 1.
Figure 1.
Figure 1.

(A) The survival of MKN45/PAR1 cells treated with paclitaxel (PTX) was assessed using the MTT assay under different concentrations and time conditions. (B) Representative images of MKN45/mock cell-bearing mice are depicted in the photographs with the drug intraperitoneally administered shown above each image and the mean of each PCI score presented immediately below. A graph of tumor weights is presented in the bottom panel. (C) Body weight and weight gain of the mice inoculated with MKN45/mock cells into the stomach wall were monitored over a period of 28 days after inoculation. Data are mean±standard deviation. (D) Survival of the MKN45/mock cell-bearing mice according to the Kaplan–Meier method is shown. PBS: Phosphate buffered saline; PAR1: protease-activated receptor-1; PCI: Peritoneal Cancer Index.

Figure 2.
Figure 2.

Graph showing the IC50 of PTX under different conditions of treatment of MKN45/PAR1 cells with different SCH79797 concentrations for 72 h. SD values are shown on the lines showing PTX (black line), PTX plus 30 nM SCH79797 (gray line), PTX plus 70 nM SCH79797 (light blue line), and PTX and 150 nM SCH79797 (dotted line). IC50: 50% Inhibitory concentration; PTX: paclitaxel; PAR1: protease-activated receptor-1; SD: standard deviation.

Next, to assess whether the treatment with SCH79797 increased drug sensitivity in vitro, the IC50 value of PTX was investigated with SCH79797 added to the culture medium. The IC50 value for PTX under this condition was 0.0145±0.009 μM (Figure 2).

SCH79797 inhibits peritoneal carcinomatosis by MKN45/PAR1 cells in nude mice. We examined MKN45/PAR1 cells, which are more likely to develop peritoneal dissemination than MKN45/mock cells, and the effect of SCH79797 on the development of experimental peritoneal carcinomatosis in mice. Mice that received MKN45/PAR1 cells in the stomach wall and were kept for 8 weeks without any treatment had a higher PCI score, which assesses the extent of peritoneal dissemination [23.8 (18-32) vs. 11.5 (10-13), p<0.001], and heavier tumor weight than mice that received MKN45/mock cells in the stomach wall and were kept for 8 weeks without any treatment (Figure 3). Both groups of mice had tumor nodules in the stomach wall.

Figure 3.
Figure 3.

Assessment of peritoneal dissemination of MKN45/PAR1 cells in mice. An MKN45/PAR1 cell-bearing mouse is depicted in the images on the left. This mouse had bloody ascites with a number of peritoneal disseminations found in the abdominal cavity. The images on the right show a MKN45/mock cell-bearing mouse. This mouse had no ascites, and a few peritoneal disseminations were found in the abdominal cavity. PCI scores for both groups are shown under the figure; the difference between groups is significant (p<0.001). The graph on the right-hand side shows the difference in tumor weights between the two mouse groups, which is also significant (p<0.001). PAR1: protease-activated receptor-1; PCI: Peritoneal Cancer Index.

In the mice inoculated with MKN45/PAR1 cells into the stomach, the PCI score and the tumor weight were smaller in the group receiving PTX and SCH79797 intraperitoneally compared to those in the PBS group [22.5 (18-30) vs. 5.2 (4-7), p<0.001; 1,086±127.2 mg vs. 33.2±19.9 mg, p<0.001], and each group that received SCH79797 or PTX had a lower PCI score and tumor weight than the PBS group [22.5 (18-30) vs. 13.8 (16-11), p=0.164; 1,086±127.2 mg vs. 487.5±50.6 mg, p=0.086 and 22.5 (18-30) vs. 9.8 (8-12), p=0.009; 1,086±127.2 mg vs. 216±86.8 mg, p=0.005] (Figure 4). In mice inoculated with MKN45/mock cells into the stomach, tumor weight was not substantially different between the group that received only SCH79797 and the untreated group, nor was tumor weight much different between the group that received only PTX and the group that received PTX and SCH79797 (Figure 1A). In mice inoculated with MKN45/mock cell into the stomach, the mean PCI scores of the PBS and SCH79797 groups were 11.5 [10-13] and 12.3 [11-14], and those of the PTX and PTX+SCH79797 groups were 8.5 [7-10] and 8 [7-9], respectively (Figure 1B).

Figure 4.
Figure 4.

Assessment of the tumor shrinkage effect of each drug on the peritoneal dissemination of MKN45/PAR1 cells. Representative images of MKN45/PAR1 cell-bearing mice are shown in the upper panel. The drug intraperitoneally administered is shown above the image for each group with the mean PCI score presented immediately below. The graphs of PCI score and tumor weights presented in the bottom panel shows the p-values for the means of each group compared with the PBS group as the reference. PCI: Peritoneal Cancer Index; PBS: phosphate buffered saline; PTX: paclitaxel; PAR1: protease-activated receptor-1.

The body weight changes of mice were evaluated up to 28 days after MKN45/PAR1 or MKN45/mock cells were inoculated into the mice stomach (Figure 1C and Figure 5). The body weight at the beginning of the experiment was not different between the groups. Body weights increased steadily in the control mice, which were not inoculated, but not in the mice inoculated with MKN/45PAR1 or MKN45/mock cells, resulting in a difference in body weight of approximately 10% regardless of tumor weight on day 14 after inoculation. Groups of mice inoculated with MKN45/PAR1 cells in the stomach and treated intraperitoneally with PTX and/or SCH79797 to inhibit tumor growth gradually gained weight two weeks after cell inoculation. However, the PBS, SCH79797, and PTX groups showed significant differences in the degree of weight gain at the end of the experimental period compared to the control group (p=0.002, p=0.013, and p=0.038), while the PTX+SCH79797 group showed no significant differences (p=0.155). In the groups of mice inoculated with MKN45/mock cells and treated intraperitoneally with PTX and/or SCH79797 to suppress tumor growth, body weight change in the group receiving SCH79797 was similar to that in the control group. Further, the group of mice receiving both SCH79797 and PTX intraperitoneally had a similar weight gain curve to the group of mice receiving PTX only intraperitoneally, with a slightly greater weight gain noted than that seen in the groups receiving a single drug injection (Figure 1C).

Figure 5.
Figure 5.

Assessment of body weight change over time in each group. Body weight and weight gain of the mice inoculated with MKN45/PAR1 cells into the stomach wall were monitored over a period of 28 days after inoculation. Data are mean±standard deviation, n=5 per group, with the p-value compared with non-inoculated mice as the control at the end of experimental period. White squares indicate the body weight of mice with no inoculated MKN45/PAR1 cells as the control; black squares indicate the body weights of mice treated with PBS after inoculation with MKN45/PAR1 cells; solid black triangles indicate the body weights of mice treated with 70 nM SCH79797 after inoculation with MKN45/PAR1 cells; white triangles indicate the body weights of mice treated with PTX after inoculation with MKN45/PAR1 cells; and black rhombuses indicate the body weights of mice treated with PTX and 70 nM SCH79797 after inoculation with MKN45/PAR1 cells. *p=0.002; **p=0.013; ***p=0.038; ****p=0.155. PBS: phosphate buffered saline; PTX: paclitaxel; PAR1: protease-activated receptor-1.

The representative results of mice at 56 days after MKN45/PAR1 cell inoculation are shown in Figure 6. The group of mice receiving PBS died within 28-35 days, resulting in a mean survival time for the PBS group of 30±3.08 days. Contrastingly, the mean survival time of the mice treated with SCH79797, PTX, and combination treatment with SCH79797 and PTX was 34.8±4.96, 45.8±6.06, and 53.6±5.37 days, respectively. Compared to the control group, the groups treated with PBS (p=0.002), with 70 nM SCH79797 (p=0.002), and with PTX (p=0.003) had significantly worse survival rates, whereas the group treated with PTX and 70 nM SCH79797 combined showed no significant difference in overall survival compared to the control group (p=0.317). The mean survival times of mice at 56 days after inoculation with MKN45/mock cells were 30.2 days in the PBS group, 31.2 days in the SCH79797 group, 49.2 days in the PTX group, and 48.5 days in the PTX+SCH79797 group (Figure 1D).

Figure 6.
Figure 6.

Survival of the MKN45/PAR1 cell-bearing mice according to the Kaplan–Meier method is shown. (A) There was a significant difference in survival rate between the PBS and control group (p=0.002). (B) There was a significant difference in survival rate between the group treated with 70 nM SCH79797 and control group (p=0.002). (C) There was a significant difference in survival rate between the PTX and control group (p=0.003). (D) There was no significant difference in survival rate between the group treated with PTX + 70 nM SCH79797 and the control group (p=0.317). p-Values were computed using log-rank test. PBS: Phosphate buffered saline; PTX: paclitaxel; PAR1: protease-activated receptor-1.

Discussion

We showed that MKN45/PAR1cells, a PAR1-positive gastric cancer cell line, was more likely to develop peritoneal dissemination than MKN45/mock cells, a PAR1-negative gastric cancer cell line, and SCH79797, a PAR1 antagonist, showed marked antitumor effects when combined with cytotoxic chemotherapy against PAR1-positive cells. These results indicated that thrombin, which increases due to a rise in the concentration of thrombin-antithrombin III complex (a degradation product of thrombin) in the ascites of patients with peritonitis (28), stimulates thrombin receptor-positive gastric cancer cells, making them prone to peritoneal metastasis in vivo and that antagonists against the thrombin receptor may be new agents for cancer therapy.

Peritoneal dissemination occurs when intraperitoneal free cancer cells leave the serous membrane of the primary tumor. For cancer cells to leave the primary site, they must break the tight adhesions between surrounding cells, free themselves, and migrate. The loss of cell adhesion and the acquisition of mesenchymal functions, such as migration and invasion, are called the epithelial-mesenchymal transition (EMT). We have shown that thrombin enhances the expression of EMT markers in PAR1-positive gastric cancer cells, promotes their invasive potential, and reduces the expression level of E-cadherin (18). The calcium-dependent cell-cell adhesion protein E-cadherin is essential for establishing epithelial architecture and maintaining cell polarity and differentiation (29, 30). It has also been shown that cancer cells have the ability to spread to distant organs and that there are significant differences between cancer cells and extracellular matrix elements (31). E-cadherin dysfunction plays a major role in the invasion of the stomach wall and the migration of cancer cells into the free abdominal space, both of which are widely implicated in the course of gastric cancer (32, 33). Furthermore, it has been reported that MMP1, an agonist of PAR1 (13), is also involved in this process and that MMP1 is produced not by gastric cancer cells but by surrounding stromal cells (34, 35).

In this study, we demonstrated in vivo that PAR1 plays an important role in the process of gastric cancer cell dissemination from the primary tumor to the peritoneum. Based on previous reports and our study, it is surmised that the important role of PAR1 is probably in the process of detachment of cancer cells from the primary tumor, during which in vivo thrombin and MMP1 produced by the stromal tissue surrounding cancer cells activate PAR1 and induce peritoneal dissemination.

The optimal treatment for peritoneal dissemination of gastric cancer has not yet been found. Patients with systemic chemotherapy are often managed in a manner similar to those with other distant metastases. The anticancer drug PTX exhibits broad antitumor effectiveness against ovarian, breast, gastric, and lung malignancies (36, 37). This medication causes cell death by stabilizing polymerized microtubules and improving microtubule assembly, which stops cells in the G0/G1 and G2/M phases of the cell cycle. Due to the large molecular weight and bulky structure of this drug, which allow it to remain in the peritoneal cavity for extended periods of time at high concentrations, intraperitoneal administration of PTX is becoming more and more popular as an effective treatment for peritoneal metastasis (38, 39). The National Cancer Institute has approved the intraperitoneal PTX regimen after it was shown to extend survival in a phase III study including ovarian cancer with peritoneal metastases (40). In patients with peritoneal metastases from gastric cancer, a recent phase II study of intravenous and intraperitoneal PTX combined with S-1 (an oral fluoropyrimidine derivative combining tegafur with two modulators) showed a 1-year overall survival rate of 78% with a median survival time of 22.5 months (41). However, in a multicenter phase III study, median survival was 17.7 months in the intraperitoneal group and 15.2 months in the standard therapy group (hazard ratio=0.72, 95% confidence interval=0.49-1.04, p=0.08), which failed to demonstrate a significant difference in the primary endpoint (42). In the present study, we showed that the IC50 value of PTX for PAR1-positive gastric cancer cells was significantly reduced by combination with a PAR1 antagonist and that mice with peritoneal dissemination of PAR1-positive gastric cancer cells in the group treated with both intraperitoneal PTX and SCH79797 had significantly fewer intraperitoneal tumor nodules and a longer prognosis. However, treatment with a PAR1 antagonist alone was not as effective, but there were fewer nodules disseminated peritoneally and prognosis was prolonged compared to that in the PBS group. When cancer cells create an environment that suppresses the immune system to evade immune cell attack, they enter a state known as immune escape (43). However, we believe that tumor cells must evade the host’s immune system, including natural killer cells, for cancer cells to appear as peritoneal recurrence. Patients with malignant ascites exhibit thrombin-like activities (44). In such an environment, thrombin significantly enhances the immunological escape strategy of tumor cells by downregulating IL-12 expression and up-regulating IL-10 expression through PAR1 (45). Additionally, the binding between autoantibodies against PAR1 and PAR1 averts cleavage and activation of the receptor by MMP1 and reduces velocity of the affected cells, as well as the number of invading cells (46). In addition, the serum level of the autoantibodies against PAR1 is significantly lower in patients with primary epithelial ovarian cancer and with high-grade carcinoma compared to healthy controls (47). In xenograft models of peritoneal dissemination, the inhibition of angiogenesis, ascites formation, and metastasis has been observed through the administration of intracellular pepducins, which disrupt PAR1 signaling (48). These results suggest that PAR1 antagonists may suppress the immune escape mechanism of cancer cells by thrombin via PAR1. Furthermore, we believe that PAR1 antagonists not only have the potential to amplify the effects of cytotoxic drugs such as PTX, but also that PAR1 has the potential to become a molecular target against peritoneal dissemination.

In conclusion, we showed that PAR1 may be a viable therapeutic target for peritoneal dissemination and that it plays a significant role in the development of peritoneal dissemination in gastric cancer. The present preclinical study suggests the need for a clinical study to evaluate the antitumor effects of PAR1 antagonists in gastric cancer patients with peritoneal metastasis.

Footnotes

  • Authors’ Contributions

    DF conceived and designed the study and performed data acquisition and statistical analysis. KH provided statistical advice and analysis assistance and prepared the manuscript. DF and KH confirm the authenticity of all of the raw data. Both authors read and approved the final manuscript.

  • Conflicts of Interest

    The Authors declare that they have no competing interests in relation to this study.

  • Funding

    This work was supported in part by the Japan Society for the Promotion of Science KAKENHI (grant no. 22K07260).

  • Received September 13, 2024.
  • Revision received October 3, 2024.
  • Accepted October 7, 2024.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY-NC-ND) 4.0 international license (https://creativecommons.org/licenses/by-nc-nd/4.0).

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PAR1 Is a Candidate Target for the Treatment of Peritoneal Dissemination in Gastric Cancer
DAISUKE FUJIMOTO, HIROTOSHI KOBAYASHI
Anticancer Research Nov 2024, 44 (11) 4857-4867; DOI: 10.21873/anticanres.17311
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