Proliferation,Migration And Invasion Effect And Mechanism Of Long Non-coding RNA LINC01088 In Ovarian Cancer Cells

Objective:To investigate the expression ofLong intergenic non-protein coding RNA 1088(LINC01088)in epithelial ovarian cancer(EOC)tissue and ovarian cancer cells;to explore the relationship between the expression of LINC01088 and the proliferation,migration,invasion of ovarian cancer cell line SKOV3;and to analyze the effect of LINC01088 on the epithelial-mesenchymal transition(EMT)of ovarian cancer cell line SKOV3 through the PI3K/Akt signaling pathway.Methods:(1)The data about theexpression levelof LINC01088 in ovarian cancer tissue and normal ovarian tissue were downloaded from the Gene Expression Profiling Interactive Analysis(GEPIA)database.(2)The expression level of LINC01088 in 24 EOC tissue samples and 24 normal ovarian tissue samples were detected by Real-time fluorescence quantitative PCR(qRT-PCR),and the relationship between LINC01088 expression level and ovarian cancer FIGO stage,lymph node metastasis of ovarian cancerwas analyzed.(3)Building LINC01088 small interference RNA(siRNA)lentivirus vectorand LINC01088 overexpression lentivirus vector,and designingthe corresponding negative control lentivirus vector,then SKOV3 cells are infected by themseparately.The stable infected cell lines and the negative control cell lines were screened out by adding the lentivirus solutions to SKOV3 cells and using the fluorescence and puromycin,qRT-PCR was using to detect LINC01088 mRNA expressionlevelin infected target cells.Four groups of experimental cells were constructed,they are the overexpression LINC01088 group(LV-LINC01088 group)and the correspondingnegative control group(LV-NC group),interference LINC01088 expression group(siRNA-LINC01088 group)and the correspondingnegative control group(siRNA-NC group).Cell proliferation wasobserved by Clone Formation assay and CCK-8 assay,migration and invasion ability were assessed by wound healing assay and Transwell assay.The growth of SKOV3 cells in siRNA-LINC01088 groupand siRNA-NC group in vivo was detected by nude mice tumorigenesis experiment.The protein levelexpression of EMT related markers E-cadherin,vimentin and PI3K/Aktsignalingpathway target gene PI3K,p-PI3K,Akt and p-Akt was detected by Western blot.(4)Ovarian cancer SKOV3 cells were treated with SC79,the PI3K/Akt signaling pathway activator,then cell proliferation was assessed using CCK-8 assay.SC79+LV-LINC01088 wasregarded as the experimental group,and SC79+LV-NC wasregarded as the negative control group.The protein expression levelof E-cadherin,vimentin p-PI3K,p-Akt was detected by Western blot.Results:(1)The GEPIA database shows that LINC01088 has expression differences between ovarian cancer tissues and normal ovarian tissues,and its expression levelis down-regulated in ovarian cancer tissues.(2)The expression level of LINC01088 mRNA was significantly lower in EOC tissue(1.16±0.74)than normal ovarian tissues(4.12±1.45)(t=13.064,P<0.01).With the progression of FIGO stages,the expression levelof LINC01088 was down-regulated(t=11.538,P<0.001),and patients with lymph node metastasis the expression level of LINC01088 mRNA was lower than patients without lymph node metastasis(t=8.401,P<0.001).Compared with normal ovarian cell line ISEO80 and ovarian cancer cell line CAOV3,the expression level ofLINC01088 mRNA is significantly lower in ovarian cancer cell line SKOV3(F=12.234,P<0.001).(3)Compared with the negative control siRNA-NC group,the proliferation,migration and invasion ability of siRNA-LINC01088 group were significantly improved.Interference LINC01088 expression can promote the growth ability of human ovarian cancer cell SKOV3 in nude mice(t=14.386,P<0.001).The expression level of E-cadherin protein was significantly decreased,however,the protein expression of Vimentin,p-Akt 473 and p-PI3K were significantly increased(P<0.01).Compared with the negative control LV-NC group,the cell proliferation,migration and invasion ability of the LV-LINC01088 group were significantly decreased.The expression levelof E-cadherin protein was increased,and the protein expression of Vimentin,p-Akt 473 and p-PI3K were significantly decreased(P<0.01).(4)SC79 can significantly promote the proliferation of SKOV3 cells,and its fittestconcentration anddurationare 10μM and 48 hours.In addition,the promote effect of each concentration is growing stabilized at 50h,72h(P>0.05).The E-cadherin protein expression level in the SC79+siRNA-LINC01088 group increased,and the vimentin,p-Akt 473 and p-PI3K protein expression level decreased significantly in contrast to the SC79+LV-NC group,and the difference was statistically significant(all P<0.01).Conclusions:(1)LINC01088 is significantly down-regulated in human epithelial ovarian cancer tissues and ovarian cancer cell lines,and its expression level is related to the patient’s FIGO stage and lymph node metastasis.(2)LINC01088 can inhibit the proliferation,migration and invasion of ovarian cancer cell SKOV3 confirmed by tests in vitro and in vivo,which means that LINC01088 may be involved in the development of ovarian cancer.(3)Overexpression of LINC01088 inhibits the EMT of ovarian cancer SKOV3 cells,and inhibition of PI3K/Akt signaling pathway may be its mechanism.Therefore,investigating the regulatory mechanism of LINC01088 in ovarian cancer may contribute to clarifing the molecular mechanism of invasion and metastasis in ovarian cancer,and providing a new marker for the diagnosis and treatment of ovarian cancer.