The Effect of RhoC siRNA on the Invasiveness and Proliferation of Human Cervical Cancer Cell Line SiHa Cells

This study investigated the effect of RhoC GTPase on the proliferation and metastasis of cervical cancer cells, SiHa cells, in vitro. RhoC siRNA was introduced into SiHa cells to silence the RhoC gene. The mRNA and protein expression of RhoC, before and after RhoC siRNA transfection, was examined by RT-PCR and Western blotting, respectively. The proliferation and apoptosis of SiHa cells were examined by MTT assay and flow cytometry （FACS）, respectively. Adhesive rate was evaluated by Matrigel adhesive assay, and the invasive capability and migration capability were assessed by transwell invasive assay and migration assay, respectively. The results showed that after the RhoC siRNA transfection, the mRNA and protein expression of RhoC was down-regulated in SiHa cells. The down-regulation of RhoC GTPase did not affect the cell proliferation and apoptosis （P〉0.05）, but it did suppress SiHa cells＇ adhesion to matrigel （P〈0.01）, the invasive capability （P〈0.01） and the migration capability （P〈0.01）. It was concluded that RhoC obviously promotes the adhesion, invasion and migration of SiHa cells in vitro, but not proliferation and apoptosis, suggesting that RhoC plays an important role in the progression in cervical cancer.

RhoC GTP酶的表达与宫颈癌侵袭和转移的关系

目的研究RhoC GTP酶的表达及其与宫颈癌侵袭和转移的关系。方法半定量RT-PCR法和免疫组化方法分别检测正常宫颈组织、宫颈上皮内瘤样变(CIN)组织和宫颈癌组织中RhoC基因mRNA和蛋白的表达;半定量RT-PCR法和Western blot分别检测人宫颈癌细胞株SiHa、HeLa和C33a中RhoC mRNA和蛋白的表达,基质胶粘附实验、Transwell体外侵袭实验和迁移实验分别比较3株宫颈癌细胞的粘附率、侵袭能力和运动能力。结果 RhoC的mRNA和蛋白表达在正常宫颈组织、CIN组织和宫颈癌组织中逐渐增高,在宫颈癌组织中的表达明显高于CIN组织;RhoC在C33a细胞中的表达明显低于SiHa和HeLa细胞,C33a细胞对细胞外基质的粘附率、体外侵袭能力和运动能力明显低于其它2种细胞。结论宫颈癌组织中,RhoC GTP酶的表达与宫颈癌的进展和淋巴结转移有关;在宫颈癌细胞中,RhoCGTP酶的表达与细胞的粘附能力、侵袭能力和运动能力呈正相关。

RhoC—siRNA对裸鼠结肠癌移植瘤的生长抑制作用

目的探讨小干扰RNA（siRNA）载体介导抑制RhoC表达对人结肠癌裸鼠移植瘤的抑制作用。方法将人结肠癌细胞HT-29接种于裸鼠背部皮下，成功建立裸鼠移植瘤模型；成瘤后将RhoC-siRNA注射入裸鼠移植瘤瘤体，观察RhoC—siRNA对裸鼠移植瘤生长的影响，Westernblot检测瘤体中RhoC、基质金属蛋白（MMP-2）和血管内皮生长因子（VEGF）蛋白表达。结果RhoC—siRNA对结肠癌裸鼠移植瘤的生长有明显抑制作用，瘤体体积重量明显减轻；RhoC—siRNA治疗后瘤体内VEGF和MMP-2蛋白表达明显下降。结论通过抑制裸鼠结肠癌移植瘤中RhoC的表达，可以抑制肿瘤的生长并下调VEGF和MMP-2的表达水平。