Effects of Targeting lncRNA HOTAIR on the Invasion and Nude Mouse Tumorigenicity of Epithelial Ovarian Cancer Cells
Abstract
To explore the effects of shRNA-mediated downregulating lncRNA HOTAIR on the invasion, nude mouse tumorigenicity and snail expression of epithelial ovarian cancer SKOV3 cells. Methods The expression of lncRNA HOTAIR was detected by RT-PCR in SKOV3 cells. The shRNA targeting the lncRNA HOTAIR gene was cloned into RNA interference plasmid. The construction shHOTAIR vector was transfected into ovarian cancer SKOV3 cells by lipofectamine 2000, and the stably transfected cells were isolated by G418 and single clone selection. The downregulating expression of lncRNA HOTAIR was detected by quantitative real time PCR(qRT-PCR). The characteristics of shHOTAIR transfected SKOV3 cells were analyzed from the assays of invasion and nude mouse tumorigenicity, as well as the expression of snail and E-cadherin mRNA detected by qRT-PCR, and snail detected by immunohistochemistry and Western blot methods in xenograft tumor, respectively. Results The lncRNA HOTAIR expression was proved by RT-PCR in SKOV3 cells. The lncRNA HOTAIR expression in shHOTAIR transfected SKOV3 cells was significantly lower than the scramble control (P<0.01). The shHOTAIR transfected SKOV3 cells show that the invasion ability was significantly decreased compared with the scramble control (P<0.01). The nude mouse tumorigenicity, including tumorigenicity mouse number and tumor volume, was significantly decreased compared with the control group (P<0.01). The snail protein expression detected by IHC and Western blot in shHOTAIR-SKOV3 xenograft tumor was significantly decreased compared with the control scramble- SKOV3 group (P<0.05). The lncRNA HOTAIR low expression resulted in increasing E-cadherin and decreasing snail expression detected by qRT-PCR in the shHOTAIR transfected SKOV3 cells of xenograft tumor, compared with the scramble control (P<0.05). Conclusion Targeting lncRNA HOTAIR expression in SKOV3 cells with RNA interference can decrease snail, increase E-cadherin and significantly reduce the invasion and tumorigenicity of epithelial ovarian cancer SKOV3 cells. These results suggest that the lncRNA HOTAIR could be an effective therapeutic target for human epithelial ovarian caner treatment.
Keywords: Epithelial ovarian cancer lncRNA, HOTAIR, RNA interference, Tumorigenicity snail
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WU H. SHANG X, SHI Y, et al. Genetic variants of IncRNA HOTAIR and risk of epithelial ovarian cancer among Chinese women . Oncotarget,2016,7(27) ;41047-41052. ZHONG Y, GAO D. HE S, et al. Dysregulated Expression of Long Noncoding RNAs in Ovarian Cancer . Int J Gynecol Cancer,2016,26(9): 1564-1570.
TESCHENDORFF AE, LEE SH, JONES A, et al. HOTAIR and its surrogate DNA methylation signature indicate carboplatin resistance in ovarian cancer. Genome Med,2015,7; 108 [2017-05-06]. https://doi. org/10. 1186/ si 3073-015-0233-4.
NIINUMA T, SUZUKI H, NOJIMA M, et al. Upregulation of miR-196a and HOTAIR drive malignant character in gastrointestinal stromal tumors. Cancer Res, 2012,72(5):1126-1136.
OZESAR, MILLER DF, OZES ON, et al. NF-кВ- HOTAIR axis links DNA damage response, chemoresistance and cellular senescence in ovarian cancer. Oncogene,2016 ,35 (41) :5350-5361.
BATTISTELLI C, CICCHINI C, SANTANGELO L, etal. The Snail repressor recruits EZH2 to specific genomic sites through the enrollment of the IncRNA HOTAIR in epithelial- to-mesenchymal transition. Oncogene,2017,36(7) :912-955.
ZHANG J. ZHANG P, WANG L. et al. Long non-coding RNA HOTAIR in carcinogenesis and metastasis. Acta Biochim Biophys Sin (Shanghai) ,2014 ,46( 1): 1-5.
YU H, SHEN Y, HONG J, et al. The contribution of TGF- |3 in epithelial-mesenchymal transition ( EMT ); down- regulation of E-cadherin via snail. Neoplasma,2015,62( 1); 1- 15.
PADUA ALVES C, FONSECA AS, MUYS BR, et al. Brief report; the lincRNA Hotair is required for epithelial-to- mesenchymal transition and sternness maintenance of cancer cell lines. Stem Cells,2013,31( 12) :2827-2832.
LUCZAK A, SUPERNAT A, LAPINSKA-SZUMCZYK S. et al. HOTAIR in relation to epithelial-mesenchymal transition and cancer stem cells in molecular subtypes of endometrial cancer. Int J Biol Markers, 2016, 31 ( 3 ); e245- e251 [ 2017-05-06 ]. http://www. biological-markers. com/ article/830badbb-8e0f-4e92-a8ce-34215598cf8a. doi: 10. 5301/ jbm. 5000187.
QIU JJ, LIN YY. YE LG, et al. Overexpression of long non-coding RNA HOTAIR predicts poor patient prognosis and promotes tumor metastasis in epithelial ovarian cancer . Gynecol Oncol, 2014,134 (1); 121-128.
DHAMIJA S, DIEDERICHS S. From junk to master regulators of invasion; IncRNA functions in migration, EMT and metastasis. Int J Cancer,2016 ,139(2): 269-280.
YEP. WANG T. LIU WH. et al. Enhancing HOTAIR/ MiR-lOb drives normal liver stem cells toward a tendency to malignant transformation through inducing epithelial- to- mesenchymal transition. Rejuvenation Res.2015.18(4)?332- 340.
DENG J. YANG M. JIANG R. et al. Long non-coding RNA HOTAIR regulates the proliferation, self-renewal capacity, tumor formation and migration of the cancer stem-like cell (CSC) subpopulation enriched from breast cancer cells. PLoS One,2017.12(1): e0170860[2017-05-06]. https://doi. org/ 10. 1371/journal, pone. 0170860.
REN K, LI Y, LU H. et al. Long noncoding RNA HOTAIR controls cell cycle by functioning as a competing endogenous RNA in esophageal squamous cell carcinoma. Transl Oncol.2016.9(6):489-497.
LAU EY, HO NP. LEE TK. Cancer stem cells and their microenvironment;biology and therapeutic implications. Stem Cells Int,2017,2017: 3714190 [2017-05-06]. https://doi. org/10. 1155/2017/371 1190.
CH1YOMARU T, YAMAMURA S, FUKUHARA S, et al. Genistein inhibits prostate cancer cell growth by targeting miR-34a and oncogenic HOTAIR . PLoS One, 2013, 8 (8) : e70372 [ 2017-05-06 ]. https;//doi. org/10. 1371/journal, pone. 0070372.
CHIYOMARU T, FUKUHARA S, SAINI S, et al. Long non-coding RNA HOTAIR is targeted and regulated by miR- 141 in human cancer cells. J Biol Chem, 2014, 289 ( 18 ) : 12550-12565.
DENG Q. SUN H, HE B, et al. Prognostic value of long non-coding RNA HOTAIR in various cancers. PLoS One. 2014,9 ( 10): el 10059 [2017-05-06]. https://doi.org/ 10. 1371/journal, pone. 0110059.
CHENG JC, CHANG HM, XIONG S, et al. Sprouty2 inhibits amphiregulin- induced down-regulation of E-cadherin and cell invasion in human ovarian cancer cells. Oncotarget, 2016,7(49);81645-81660.
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