Heat shock protein 70-2 (HSP70-2) a novel cancer testis antigen that promotes growth of ovarian cancer

Gupta, N and Jagadish, N and Surolia, A and Suri, A (2017) Heat shock protein 70-2 (HSP70-2) a novel cancer testis antigen that promotes growth of ovarian cancer. In: American Journal of Cancer Research, 7 (6). pp. 1252-1269.

PDF ame_jou_can_1252-1269_2017.pdf - Published Version Restricted to Registered users only Download (3MB) | Request a copy

Abstract

Heat shock protein 70-2 (HSP70-2) is known to be involved in tumor progression. However, its molecular role and mechanism in epithelial ovarian cancer (EOC) remains unknown. In the present investigation, we examined the role of HSP70-2 in cell cycle, apoptosis and epithelial mesenchymal transition pathways in EOC cells in in vitro and in-vivo xenograft mouse model. To investigate the role of HSP70-2 in ovarian cancer, plasmid driven short hairpin RNA approach was used to examine HSP70-2 gene and protein expression in ovarian cancer cell line A-10 (origin: serous papillary cystadenocarcinoma), Caov-3 (origin: adenocarcinoma) and SKOV3 (origin: adenocarcinoma; derived from metastatic site: ascites) by RT-PCR, quantitative-PCR, immunohistochemistry and Western blotting. Light microscopy, scanning electron microscopy, viability tests, and flow cytometry were used to study the cellular proliferation, onset of senescence, colony forming ability and morphological features of cancer cells. Cell migration and invasion ability was evaluated by wound healing and Boyden chamber assays. Further, we studied the effect of HSP70-2 protein ablation on human ovarian xenograft mice model. At molecular level, various molecules involved in apoptosis, cell cycle and epithelial-mesenchymal-transition were also examined both in in-vitro and in-vivo xenograft mouse model. The knockdown of HSP70-2 expression by gene silencing resulted in the onset of apoptosis, senescence, reduced cellular growth and colony forming ability of EOC cells. Interestingly, the migration, invasion and wound healing abilities of cells were also significantly inhibited. In addition, the ablation of HSP70-2 resulted in the upregulation of cytochrome-C, caspase 3, caspase 7, caspase 9, APAF1, BAX, BIM, BAK, BAD, BID, PUMA, NOXA, p16, p21, Rb, E-cadherin, cytokeratin 18, EMA in these cells as well as in the xenograft tumor specimens. However, there was downregulation of PARP1, BCL-2, Bcl-xL, MCL-1, Survivin, XIAP, cIAP2, CDK1, CDK2, CDK4, CDK6, cyclin D1, cyclin E, cyclin A2, cyclin B1, p-Rb, N-cadherin, SNAIL, SLUG, VIMENTIN, SMA, MMP2, MMP3, MMP9 and TWIST in these samples. Furthermore, the xenograft studies showed significant reduction in the tumor growth. Our results suggest that HSP70-2 can promote cellular growth and invasion of EOC cells and therefore may be a potential therapeutic target in EOC.

Item Type:	Journal Article
Publication:	American Journal of Cancer Research
Publisher:	E-Century Publishing Corporation
Additional Information:	The copyright for this article belongs to E-Century Publishing Corporation
Keywords:	apoptotic protease activating factor 1; BIM protein; cancer testis antigen; caspase 3; caspase 7; caspase 9; cyclin dependent kinase 1; cyclin dependent kinase 2; cytochrome c; cytokeratin 18; heat shock protein 70; heat shock protein 70-2; nicotinamide adenine dinucleotide adenosine diphosphate ribosyltransferase 1; protein BAD; protein Bak; protein Bax; protein bcl 2; protein bcl xl; protein Bid; protein mcl 1; protein Noxa; protein p16; protein p21; PUMA protein; retinoblastoma protein; short hairpin RNA; survivin; unclassified drug; uvomorulin; X linked inhibitor of apoptosis, animal experiment; animal model; animal tissue; apoptosis; Article; carcinogenesis; cell aging; cell cycle; cell growth; cell invasion; cell migration; cell proliferation; cell structure; cell viability; colony formation; controlled study; down regulation; epithelial mesenchymal transition; female; flow cytometry; gene expression; gene silencing; human; human cell; immunohistochemistry; in vitro study; in vivo study; microscopy; mouse; nonhuman; ovarian cancer cell line; ovary cancer; polymerase chain reaction; protein expression; quantitative analysis; reverse transcription polymerase chain reaction; scanning electron microscopy; tumor xenograft; upregulation; Western blotting; wound healing assay
Department/Centre:	Division of Biological Sciences > Molecular Biophysics Unit
Date Deposited:	18 Jul 2022 10:48
Last Modified:	18 Jul 2022 10:48
URI:	https://eprints.iisc.ac.in/id/eprint/74773

Actions (login required)

View Item