lncRNA TINCR对滋养层HTR-8/SVneo细胞生物学行为的影响及其机制

Effect of lncRNA TINCR on the biological behavior of trophoblast HTR-8/SVneo cells and its mechanism

目的探讨lncRNA组织分化诱导非蛋白编码RNA(TINCR)对滋养层HTR-8/SVneo细胞增殖、迁移、侵袭、上皮间质转化和凋亡的影响及其机制。方法将HTR-8/SVneo细胞进行体外培养,以正常培养细胞为对照,转染NC-siRNA为阴性对照,转染TINCR-siRNA干扰TNICR的表达,采用实时荧光定量PCR(qPCR)检测细胞中TINCR表达,CCK-8法检测细胞增殖,划伤愈合实验检测HTR-8/SVneo细胞迁移,Transwell小室检测细胞迁移和侵袭,Western blot检测细胞中上皮间质转化相关蛋白E-cadherin、N-cadherin、Vimentin和Wnt/β-catenin信号通路关键蛋白Wnt5a、β-catenin表达。两组间比较采用独立样本t检验,多组间比较采用单因素方差分析,组间两两比较采用SNK-q检验。结果与对照及阴性对照比较,转染TINCR-siRNA细胞后TINCR表达水平(0.25±0.03比1.00±0.08;0.25±0.03比0.98±0.06)降低;与对照及阴性对照比较,转染TINCR-siRNA细胞后HTR-8/SVneo细胞中Vimentin(0.92±0.08比0.33±0.05,0.92±0.08比0.31±0.04)、N-cadherin蛋白表达水平(0.87±0.07比0.29±0.04,0.87±0.07比0.32±0.05)均升高,而E-cadherin蛋白表达水平降低;与对照及阴性对照比较,转染TINCR-siRNA细胞后Wnt5a(0.97±0.06比0.48±0.04,0.97±0.06比0.46±0.03)、β-catenin蛋白(0.89±0.05比0.37±0.03,0.89±0.05比0.39±0.03)均升高(P均<0.05)。结论下调TINCR表达可能通过激活Wnt/β-catenin信号通路,促进HTR-8/SVneo细胞增殖、迁移、侵袭和上皮间质转化并抑制其凋亡。

Objective To investigate the effect of lncRNA tissue differentiation-inducing differentiation inducing non-protein coding RNA(TINCR)on proliferation,migration,invasion,epithelial-stromal transformation and apoptosis of trophoblast HTR-8/SVneo cells.Methods HTR-8/SVneo cells were cultured in vitro and then divided into three groups,which included the control group(standard culture),negative group(transfected NC-siRNA)and interference group(transfected TINCR-siRNA)respectively.The expression level of TINCR in the cells was detected by real-time fluorescence quantitative PCR,cell proliferation was detected by CCK-8 method,the migration of HTR-8/SVneo cells was detected by scratch healing assay,and cell migration and cell invasion were tested by Transwell chamber.The expression of epithelial-mesenchymal transformation(EMT)related proteins E-cadherin,N-cadherin,Vimentin and Wnt/β-catenin key proteins Wnt5a andβ-catenin in cells were checked by Western blotting.Results Compared with the control group and the negative group,the expression level of TINCR(0.25±0.03 vs 1.00±0.08;0.25±0.03 vs 0.98±0.06)was decreased in the interference group;compared with the control group and negative group,the expression levels of Vimentin(0.92±0.08 vs 0.33±0.05,0.92±0.08 vs 0.31±0.04)and N-cadherin(0.87±0.07 vs 0.29±0.04).(0.87±0.07 vs 0.32±0.05)were increased in HTR-8/SVneo cells in interference group,while the expression level of E-cadherin protein were decreased;compared with the control group and negative group,the expression level of Wnt5a(0.97±0.06 vs 0.48±0.04,0.97±0.06 vs 0.46±0.03)andβ-catenin proteins(0.89±0.05 vs 0.37±0.03,0.89±0.05 vs 0.39±0.03)were increased in interference group(P<0.05).Conclusion Down-regulating TINCR expression may promote the proliferation,migration,invasion and epithelial-mesenchymal transformation of HTR-8/SVneo cells and inhibit their apoptosis by activating the Wnt/β-catenin signaling pathway.