芬太尼调控LINC00184抑制卵巢癌SKOV3细胞增殖和侵袭

目的:研究芬太尼对卵巢癌SKOV3细胞增殖、凋亡和侵袭等生物学行为的影响，并探讨其作用机制。方法:体外培养人卵巢癌SKOV3细胞，采用CCK-8实验、克隆形成实验检测芬太尼对SKOV3细胞体外增殖的影响；分别采用流式细胞术和Transwell实验检测芬太尼对SKOV3细胞凋亡和侵袭能力的影响；Western blot检测细胞增殖、凋亡及侵袭相关蛋白表达。qRT-PCR检测LINC00184在卵巢癌细胞系和正常卵巢上皮细胞株的表达；构建LINC00184过表达质粒并转染SKOV3细胞，观察上调LINC00184表达对芬太尼处理的SKOV3细胞增殖、凋亡及侵袭的影响。结果:芬太尼在体外呈时间和浓度依赖性抑制SKOV3细胞存活，而不明显影响正常卵巢上皮细胞HOSE的存活率。相比对照组，使用5 ng/mL和10 ng/mL芬太尼处理SKOV3细胞，显著下调LINC00184表达水平，降低体外克隆形成率，抑制Ki67、PCNA蛋白表达（均P＜0.01）；显著增加SKOV3细胞凋亡率，上调Bax蛋白表达，下调Bcl-2蛋白表达（均P＜0.01）；显著降低侵袭细胞数，上调E-cadherin蛋白表达，下调N-cadherin、MMP9蛋白表达（均P＜0.01）。而使用LINC00184过表达质粒上调LINC00184表达则明显削弱芬太尼对SKOV3细胞的上述作用（P＜0.01）。结论:LINC00184在卵巢癌细胞表达上调，并与卵巢癌细胞的增殖、凋亡和侵袭等生物学特性相关；芬太尼通过下调LINC00184表达抑制卵巢癌细胞的增殖和侵袭，并诱导其凋亡。

Fentanyl inhibits proliferation and invasion of ovarian cancer SKOV3 cells by regulating LINC00184

Objective:To study the effects of fentanyl on the biological behaviors (proliferation,apoptosis,invasion) of ovarian cancer SKOV3 cells,and to explore its mechanism of action.Methods:Human ovarian cancer SKOV3 cells were cultured in vitro.The effects of fentanyl on proliferation of SKOV3 cells in vitro were detected by CCK-8 and clone formation assay.The effects of fentanyl on apoptosis and invasion of SKOV3 cells were detected by flow cytometry and Transwell assay.The expressions of cells proliferation,apoptosis and invasion related proteins were detected by Western blot.The expression of long intergenic non-coding RNA 00184 (LINC00184) in ovarian cancer cell lines and normal ovarian epithelial cell lines was detected by qRT-PCR.The plasmids with LINC00184 overexpression were constructed and transfected into SKOV3 cells.The effects of fentanyl on the proliferation,apoptosis and invasion of SKOV3 cells by up-regulating LINC00184 were observed.Results:Fentanyl could inhibit the survival of SKOV3 cells in vitro,showing time and concentration dependence,but could not significantly affect the survival rate of HOSE in normal ovarian epithelial cells.Compared with control group,5 ng/mL and 10 ng/mL fentanyl could significantly down-regulate the expression level of LINC00184,reduce formation rate of in vitro clone,and inhibit the expressions of Ki67 and PCNA (P＜0.01).It could significantly increase apoptosis rate of SKOV3 cells,up-regulate Bax protein and down-regulate Bcl-2 protein (P＜0.01).The fentanyl could significantly reduce number of invasion cells,up-regulate E-cadherin protein,and down-regulate N-cadherin and MMP9 proteins (P＜0.01).The application of plasmids with LINC00184 overexpression to up-regulate LINC00184 expression could significantly weaken the above-mentioned effects of fentanyl on SKOV3 cells (P＜0.01).Conclusion:The expression of LINC00184 is up-regulated in ovarian cancer cells,which is related to biological characteristics (proliferation,apoptosis,invasion) of ovarian cancer cells.Fentanyl could inhibit the proliferation and invasion of ovarian cancer cells and induce their apoptosis by down-regulating LINC00184 expression.