Oncology/Endocrine
A potential role for resveratrol as a radiation sensitizer for melanoma treatment

https://doi.org/10.1016/j.jss.2013.02.037Get rights and content

Abstract

Background

Radiotherapy (XRT) is used to improve local control of melanoma and for palliation of metastatic disease. Clinical use of XRT for melanoma is often limited by extent of disease and the relative radioresistance of melanoma may limit the effectiveness of XRT. Our group and others have previously shown that resveratrol (RSV) enhances radiation sensitivity in radioresistant prostate cancer cell lines.

Material and Methods

In this study, the effects of XRT in combination with RSV on radioresistant melanoma lines, SK-Mel-5 and HTB-65, were evaluated by assessment of proliferation and apoptosis. Clonogenic assay, comparison of proliferating cell nuclear antigen staining, Quick Cell Proliferation assay, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining and caspase-3 activity assay were used to assess proliferation and apoptosis, as appropriate.

Results

We found that the percentage of colonies, proliferating cell nuclear antigen + cells and the optical density value of melanoma cells were decreased after addition of RSV to XRT (XRT/RSV). TUNEL + cells and the relative caspase-3 activity in melanoma cells were increased after addition of RSV to XRT (XRT/RSV). We investigated the possible molecular mechanisms of decreased proliferation and increased apoptosis by using reverse transcriptase-polymerase chain reaction and immunohistochemical staining. The anti-proliferative effect of XRT/RSV correlated with decreased expression of pro-proliferative molecule cyclin B, cyclin D, cdk2 and cdk4. The pro-apoptotic effect of XRT/RSV correlated with decreased expression of the anti-apoptotic molecule FLIP, Bcl-2, and survivin.

Conclusion

These data suggest that RSV enhances radiation sensitivity of melanoma cells by inhibiting proliferation and promoting apoptosis. Resveratrol may have a potential role as a radiation sensitizer for melanoma treatment.

Introduction

Melanoma has the fastest growing incidence of any cancer among men and the second fastest growing incidence in women [1], [2], [3]. About 70,230 new cases of melanoma were projected to occur in the US in 2011 [2]. Even with recent therapeutic breakthroughs [4], [5], the management of advanced melanoma is very challenging, in part because there are few effective treatment modalities [6], [7], [8]. High dose radiotherapy (XRT) has a small but important role in the palliative management of brain metastasis, spinal cord compression, and symptomatic bony metastasis of melanoma [9], [10], [11]. A safe and effective radiosensitizing agent may allow a decrease in the radiation dose and side effects associated with XRT for advanced melanoma.

Resveratrol (RSV), a polyphenolic compound, is found in grapes, peanuts, and other plant species [12], [13], [14]. Through its complex functions, it exerts neuroprotective, immunomodulatory, anti-inflammatory, antioxidant, and antitumor functions. In recent years, RSV has been recognized as a promising anti-cancer agent [12]. Its antitumor functions have been investigated in breast cancer [15], thyroid cancer [16], squamous cell carcinoma [17], HL-60 leukemia [18], colon cancer [19], ovarian carcinoma, and prostate cancer cell lines [20], [21]. Despite widespread investigation into the direct antitumor effect of RSV, there is little data regarding the effect of RSV as a radiosensitizer for antitumor therapy.

We have recently shown that RSV enhances the effectiveness of XRT in prostate cancer, suggesting its potential role as a radiation sensitizer in other cancers [22], [23]. In this study, we studied the effect of XRT in combination with RSV on proliferation and apoptosis in a radioresistant melanoma cell lines, SK-Mel-5 and HTB-65, and investigated the potential molecular mechanisms.

Section snippets

Tumor cell line

Human SK-Mel-5 melanoma cells were provided by Thomas P. Quinn (University of Missouri, Columbia, MO). Human HTB-65 melanoma cells were purchased from ATCC (Manassas, VA). Cells were maintained in Dulbecco’s modified eagle medium (Invitrogen, Grand Island, NY), supplemented with 7% heat-inactivated fetal bovine serum and 1% penicillin-streptomycin (Invitrogen). Cells were cultured under conditions as previously described [22], [23]. Cells were grown until they reached 70% confluence, at which

Synergistic effect of RSV/XRT on inhibition of melanoma cell proliferation

Our previous studies indicated that RSV synergized with XRT to inhibit the proliferation of prostate cancer cells [22], [23]. To investigate if this synergistic effect of RSV/XRT is applicable to melanoma cells, 70% confluent SK-Mel-5 cells were treated with RSV at variable concentration (0–50 μM) for 24 h, followed by XRT at variable doses as described in the Material and Methods section. Twenty-four hours after XRT, cell survival was evaluated by clonogenic survival assay. In the absence of

Discussion

Improving the efficacy of XRT and decreasing its potential side effects might allow expanded use of XRT in the treatment to many cancers, including melanoma. We have previously shown that RSV enhances radiation sensitivity in radioresistant prostate cancer cell lines [22], [23]. In this study, we ask if RSV could synergize with XRT to inhibit growth of melanoma cells. We demonstrated that RSV synergized with XRT to inhibit proliferation and increase apoptosis of melanoma cells. The

Acknowledgment

The authors thank Dr. Thomas P. Quinn for providing SK-Mel-5 cells. They thank Dr. Xuhui Chen, Chelsea L. Ledgewood, and Abdimalik O. Mohamud for their technical assistance.

The authors declare that they have no conflict of interest.

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