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Oncol Lett. 2015 Sep;10(3):1378-1384. Epub 2015 Jun 16.

Overexpression of short TRPM8 variant α promotes cell migration and invasion, and decreases starvation-induced apoptosis in prostate cancer LNCaP cells.

Author information

1
Department of Urology, The Second Xiangya Hospital of Central South University, Changsha, Hunan 410011, P.R. China.
2
Department of Dermatology, The Second Xiangya Hospital of Central South University, Changsha, Hunan 410011, P.R. China.
3
Department of Urology, Zhongnan Hospital of Wuhan University, Wuhan, Hubei 430071, P.R. China.

Abstract

The aim of the present study was to investigate the function of a transient receptor potential melastatin 8 (TRPM8) splice variant, short TRMP8α (sM8α), in the androgen-dependent prostate cancer LNCaP cell line, and to evaluate the potential involvement of the mitogen-activated protein kinase (MAPK) signaling pathway. The coding DNA for sM8α was cloned and transfected into LNCaP cells to generate cells that overexpress this isoform of TRPM8. Cellular proliferation was determined by performing an MTT assay, and flow cytometry was used to analyze apoptosis and cell cycle distribution. Furthermore, cellular migration and invasion were evaluated using Transwell® migration assays. The subcellular location of recombinant sM8α was detected by quantum dots-based immunofluorescent imaging, western blotting was performed to examine the expression levels of proteins in the MAPK signaling pathway and reverse transcription-polymerase chain reaction was used to determine the expression of sM8α mRNA transcripts. The present study demonstrated that sM8α mRNA was expressed at a low level in the LNCaP, DU145 and PC-3 prostate cancer cell lines. Additionally, the recombinant sM8α protein was located in the cytoplasm of LNCaP cells and its overexpression significantly reduced starvation-induced apoptosis in these cells (P<0.05), possibly by means of reduced activation of phosphorylated-c-Jun N-terminal kinase (p-JNK). The migration and invasion of the LNCaP cells were markedly enhanced by the overexpression of sM8α, possibly via activation of MMP-2. Furthermore, overexpression of sM8α in LNCaP cells did not alter the expression of full-length TRPM8 and had no effect on cellular proliferation. Overall, the results of the present study indicate that sM8α may be important in the regulation of prostate cancer cell migration and invasion through the activation of matrix metalloproteinase-2, as well as in the regulation of apoptosis through the activation of p-JNK in the MAPK signaling pathway.

KEYWORDS:

alternative splicing; mitogen-activated protein kinase signaling pathway; prostate cancer; transient receptor potential melastatin 8; variant

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