TUSC3在宫颈癌组织中的表达及其对宫颈癌细胞上皮间质转化、迁移和侵袭的影响

Expression of TUSC3 in cervical cancer tissues and its effects on the epithelial-mesenchymal transition,migration and invasion of cervical cancer cells

目的探讨TUSC3在宫颈癌组织中的表达及其对宫颈癌细胞上皮间质转化、迁移和侵袭的影响。方法采用实时荧光定量PCR(qRT-PCR)检测宫颈癌组织中的TUSC3表达水平,分析其表达与宫颈癌临床病理特征的关系。构建TUSC3过表达的慢病毒载体(LV11-TUSC3)及对照载体(LV11-NC),选取过表达TUSC3的SiHa细胞作为LV11-TUSC3组,感染对照载体的SiHa细胞作为LV11-NC组,未感染慢病毒的SiHa细胞作为Con组。采用qRT-PCR检测各组细胞中TUSC3的mRNA表达水平,蛋白质免疫印迹法检测TUSC3、E-cadherin、Ncadherin、GRP78、ATF6的蛋白表达水平,MTT实验检测细胞增殖情况,划痕实验检测细胞迁移能力,Transwell实验检测细胞侵袭能力。结果宫颈癌组织中TUSC3 mRNA表达水平显著低于癌旁组织(P<0.05)。TUSC3表达水平与宫颈癌患者的年龄、肿瘤体积、宫颈浸润深度无关(P>0.05),而与FIGO分期、淋巴结转移有关(P<0.05)。与Con组和LV11-NC组相比较,LV11-TUSC3组细胞TUSC3 mRNA和蛋白表达水平升高,E-cadherin表达上调,N-cadherin表达下调,细胞增殖、迁移和侵袭能力降低,内质网应激水平降低,差异均有统计学意义(P<0.05)。结论TUSC3在宫颈癌组织中表达下调,上调其表达可抑制人宫颈癌SiHa细胞的增殖、上皮间质转化、迁移和侵袭、内质网应激。TUSC3可能成为宫颈癌治疗的潜在靶点。

Objective To explore the expression of TUSC3 in cervical cancer tissues and its effects on the epithelialmesenchymal transition,migration and invasion of cervical cancer cells.Methods Quantitative real-time PCR(qRT-PCR)was used to detect the expression level of TUSC3 in cervical cancer tissue,and the relationship between its expression and clinicopathological characteristics was analyzed.TUSC3 overexpressing lentiviral vector(LV11-TUSC3)and control vector(LV11-NC)were constructed.SiHa cells overexpressing TUSC3 were selected as the LV11-TUSC3 group,SiHa cells in‑fected with the control vector as the LV11-NC group,and those not infected with the lentivirus as the Con group.qRT-PCR was used to detect the mRNA expression of TUSC3.Western blotting was used to detect the protein expression of TUSC3,E-cadherin,N-cadherin,GRP78 and ATF6.MTT test was used to detect the proliferation of SiHa cells.Scratch test was used to detect the migration ability of SiHa cells,and Transwell test was used to detect the invasion ability of SiHa cells.Results The expression level of TUSC3 mRNA was lower in cervical cancer tissue than in tumor-adjacent tissue(P<0.05).The expression level of TUSC3 had no relation with the age,tumor volume and depth of cervical infiltration of patients(P>0.05),but was related to the FIGO stage and lymph node metastasis of patients(P<0.05).Compared with Con group and LV11-NC group,the mRNA and protein expressions of TUSC3 in cells of LV11-TUSC3 group were increased,the expres‑sion of E-cadherin was up-regulated,the expression of N-cadherin was down-regulated,the cell proliferation,migration and invasion were reduced,and the level of endoplasmic reticulum stress was decreased(P<0.05).Conclusion The expres‑sion of TUSC3 was down-regulated in cervical cancer tissues.Up-regulation of TUSC3 expression can inhibit the prolifera‑tion,migration,invasion and endoplasmic reticulum stress of human cervical cancer SiHa cells.TUSC3 may become a po‑tential target for cervical cancer treatment.