干扰TBL1XR1表达对口腔鳞癌细胞增殖和迁移的影响及其机制

Effect of disturbed expression of TBL1XR1 on proliferation and metastasis of oral squamous cell carcinoma and its mechanism

目的探讨干扰转导素β1X连锁受体蛋白1(TBL1XR1)表达对口腔鳞癌(OSCC)细胞增殖和迁移的影响及其机制。方法取OSCC细胞Cal-27,常规培养、传代。取传2代、对数生长期、生长状态良好的Cal-27细胞,随机分为无关序列组、TBL1XR1干扰组,分别转染TBL1XR1 shRNA、scramble shRNA,采用RT-qPCR法鉴定转染效率。收集两组转染72 h细胞,采用MTS法检测细胞增殖活性,采用平板克隆实验检测细胞增殖能力,采用Transwell迁移实验检测细胞迁移能力,采用Western blotting法检测JAK2/STAT3信号通路关键蛋白表达。结果TBL1XR1干扰组TBL1XR1 mRNA相对表达量低于无关序列组(t=30.53,P<0.05)。转染72 h细胞后,TBL1XR1干扰组再培养48、72、96 h时细胞增殖活性均低于无关序列组(P均<0.05),但再培养24 h时细胞增殖活性与无关序列组比较P>0.05。TBL1XR1干扰组细胞增殖和迁移能力均低于无关序列组(t分别为18.74、20.57,P均<0.05)。TBL1XR1干扰组JAK2/STAT3信号通路关键蛋白p-JAK2、p-STAT3相对表达量均低于无关序列组(P均<0.05),而JAK2、STAT3相对表达量比较P均>0.05。结论干扰TBL1XR1表达可能通过调控JAK2/STAT3信号通路关键蛋白表达,抑制OSCC细胞增殖和迁移。

Objective To investigate the effects and possible mechanisms of transducin(β)-like 1 X-linked receptor 1(TBL1XR1)on the proliferation and metastasis of oral squamous cell carcinoma(OSCC)cells.Methods OSCC cell line Cal-27 was routinely cultured and passaged.We selected the Cal-27 cells in the logarithmic phase of the second generation,and in good condition to divide them into the scramble group and sh-TBL1XR1 group.The expression of TBL1XR1 in each group was detected by RT-qPCR.The proliferation of each group was detected by MTS.The ability of cell clone formation of each group was detected by plate cloning assay.The metastasis ability of each group was detected by Transwell assay.The effect of TBL1XR1 on JAK2/STAT3 signal pathway was detected by Western blotting.Results The expression of TBL1XR1 was significantly lower in the sh-TBL1XR1 group than in the scramble group(t=30.53,P<0.05).After 72-hour transfection,the cell proliferation activities of the sh-TBL1XR1 group at 48,72,and 96 h after culture were lower than those of the scramble group(all P<0.05),but no significant difference was found in the the cell proliferation activity between these two groups at 24 h after culture(P>0.05).The cell proliferation and metastasis abilities of the sh-TBL1XR1 group were lower than those of the scramble group(t=18.74,20.57,all P<0.05).The relative expression levels of key proteins p-JAK2 and p-STAT3 of JAK2/STAT3 signaling pathway in the sh-TBL1XR1 group were lower than those of the scramble group(all P<0.05).However,there was no statistically significant difference in the relative expression of p-JAK2 and p-STAT3 protein between these two groups(P>0.05).Conclusion Interfering with the expression of TBL1XR1 may inhibit the proliferation and metastasis of OSCC cells by regulating JAK2/STAT3 signaling pathway.