RhoC siRNA对宫颈癌SiHa细胞增殖和侵袭能力的影响

目的:探讨Ras相似物C(RashomologueC,RhoC)GTP酶对宫颈癌SiHa细胞增殖和转移的影响。方法:以人宫颈癌细胞株SiHa为研究对象,利用RhoC小干扰RNA对RhoC基因进行沉默,RT-PCR和WesternBlot检测转染前后RhoC的表达变化,MTT法比较细胞增殖能力的改变,流式细胞仪检测细胞凋亡率的改变。基质胶粘附实验比较细胞粘附率的变化,transwell体外侵袭实验比较细胞侵袭能力变化,体外迁移实验比较细胞运动能力的改变。结果:RhoCsiRNA明显下调RhoC基因的表达。RhoC表达下调后,SiHa细胞的增殖能力和凋亡率没有明显变化(P>0.05),SiHa细胞对基质胶的粘附率明显降低(P<0.01),SiHa细胞的体外侵袭能力明显降低(P<0.01),体外迁移能力显著降低(P<0.01)。结论:RhoC明显地促进了宫颈癌SiHa细胞株的体外粘附、侵袭和迁移,但对其增殖和凋亡没有显著作用。提示RhoC在宫颈癌的恶性进展中起重要作用。

The inhibition effect of RhoC siRNA on invasion and proliferation of human cervical cell line SiHa

Objective：To study the effect of RhoC GTPase on proliferation and metastasis of cervical cancer cell line SiHa in vitro. Methods： RhoC siRNA was introduced into SiHa cells to silence RhoC gene. The mRNA and protein expressions of RhoC, before and after siRNA transfection, were examined by RT-PCR and Western blot, respectively. At the same time, cellular proliferation and apoptosis of cells were examined by MTT assay and flow cytometry （FACS） respectively. Adhesive rate was evaluated by Marigel adhesive assay, and the invasive capability and migration capability were evaluated by transwell invasive assay and migration assay, respectively. Results： RhoC siRNA transfection specifically down-regulated the RhoC mRNA and its protein in SiHa cells. The downregulation of RhoC GTPase did not affect the cellular proliferation and apoptotic rate of SiHa cells （P 〉0.05 ） ,but it did suppress SiHa cells＇adhesive rate to matrigel（P 〈0.01 ） ,invasive capability（P 〈 0.01 ） and migration capability（P 〈0.01 ）. Conclusion：RhoC obviously promotes the adhesion, invasion and migration of SiHa cells in vitro, but has no influence on proliferation and apoptosis. These results suggest that RhoC may play an important role in the malignant progression in cervical cancer.