Down-regulation of survivin by oxaliplatin diminishes radioresistance of head and neck squamous carcinoma cells

doi:10.1016/j.radonc.2010.06.005

Radiotherapy and Oncology

Volume 96, Issue 2, August 2010, Pages 267-273

https://doi.org/10.1016/j.radonc.2010.06.005 Get rights and content

Abstract

Background

Oxaliplatin is integrated in treatment strategies against a variety of cancers including radiation protocols. Herein, as a new strategy we tested feasibility and rationale of oxaliplatin in combination with radiation to control proliferation of head and neck squamous cell carcinoma (HNSCC) cells and discussed survivin-related signaling and apoptosis induction.

Methods

Cytotoxicity and apoptosis induced by radiation and/or oxaliplatin were examined in relation to survivin status using two HNSCC cell lines viz., Cal27 and NT8e, and one normal 293-cell line. Survivin gene knockdown by siRNA was also tested in relevance to oxaliplatin-mediated radiosensitization effects.

Results

Survivin plays a critical role in mediating radiation-resistance in part through suppression of apoptosis via a caspase-dependent mechanism. Oxaliplatin treatment significantly decreased expression of survivin in cancer cells within 24–72 h. Apoptotic cells and caspase-3 activity were increased parallely with decrease in cell viability, if irradiated during this sensitive period. The cytotoxicity of oxaliplatin and radiation combination was greater than additive. Survivin gene knockdown experiments have demonstrated the role of survivin in radiosensitization of cancer cells mediated by oxaliplatin.

Conclusions

Higher expression of survivin is a critical factor for radioresistance in HNSCC cell lines. Pre-treatment of cancer cells with oxaliplatin significantly increased the radiosensitivity through induction of apoptosis by potently inhibiting survivin.

Section snippets

Cell culture

Two HNSCC (NT8e, Cal27) and one normal 293-cell line were maintained in culture media (DMEM) with 2 mM glutamine, 10% heat-inactivated fetal bovine serum, 100 U/ml penicillin and 100 μg/ml streptomycin as per standard culture conditions. Two cell lines (NT8e and 293) were provided by Dr. Mulherkar as a gift and Cal27 cell line was obtained from American Type Culture Collection (Manassas, VA). NT8e and Cal27, shown to resist conventional cancer therapies, have primarily been isolated from upper

Radiation treatment induced survivin expression

Both cancer cell lines were highly expressive for survivin but normal 293-cell line was found negative for survivin expression (Fig. 1A). Further we examined the effect of radiation treatment on survivin expression in our selected cell lines. Radiation treatment significantly induced the expression of survivin in cancer cell lines at both doses (3 and 6 Gy) and time points (12 and 24 h) whereas no effect was detected in 293-cell line (Fig. 1B).

Over-expression of survivin provides resistance to radiation

To elucidate whether survivin is directly involved in

Discussion

Radiotherapy and surgery are the cornerstones in treating head and neck cancer [23]. Locoregional recurrence remains the major pattern of treatment failure. About 40% of patients experienced locoregional tumor progression within five years after therapy [24]. Also resistance hampers the long-term effectiveness of radiotherapy in HNSCC. Thus understanding of the molecular mechanisms contributing to drug resistance may provide opportunity to develop more effective anti-cancer therapeutic

Acknowledgment

The authors are thankful to the Council of Scientific and Industrial Research (CSIR), New Delhi for the award of Emeritus Scientist to Prof. P.S. Bisen and to Dr. Rita Mulherkar, ACRTEC, Tata Memorial Centre, Navi Mumbai for providing cell lines for this study.

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Knockdown of survivin significantly suppressed HNSCC cell proliferation and induced apoptosis in vitro. Survivin inhibition could also significantly reduce in vitro cell migration and matrigel invasion that might be due to inactivation of matrix metalloproteinases. In vivo studies showed significant repression of Cal27 xenograft tumor growth and tissue metastasis leading to improvement in mice survival in the Svv-Lent treated group compared to controls. Our data indicated that survivin expression in HNSCC cells contributed to chemo-radioresistance, and its down-regulation increased anti-cancer effects of paclitaxel, cisplatin and radiation.
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RadiobiologyDown-regulation of survivin by oxaliplatin diminishes radioresistance of head and neck squamous carcinoma cells

Abstract

Background

Methods

Results

Conclusions

Section snippets

Cell culture

Radiation treatment induced survivin expression

Over-expression of survivin provides resistance to radiation

Discussion

Acknowledgment

J Invest Dermatol

Am J Pathol

Int J Radiat Oncol Biol Phys

Int J Radiat Oncol Biol Phys

Cancer Lett

Mutat Res

Int J Radiat Oncol Biol Phys

Exp Mol Pathol

Cancer Lett

J Invest Dermatol

J Biol Chem

Head and neck cancer

N Engl J Med

Induction followed with concurrent chemoradiotherapy in advanced head & neck cancer

J Cancer Res Ther

Markers of radioresistance in squamous cell carcinomas of the head and neck: a clinicopathologic and immunohistochemical study

J Clin Oncol

Development of novel radioresistant oral squamous cell carcinoma cell lines

Clin Otolaryngol

Induction of apoptosis and inhibition of cell proliferation by survivin gene targeting

J Biol Chem

Control of apoptosis and mitotic spindle checkpoint by surviving

Nature

Radiobiology
Down-regulation of survivin by oxaliplatin diminishes radioresistance of head and neck squamous carcinoma cells