氯离子通道蛋白-3对宫颈癌细胞生长、转移的影响

Effects of chloride channel-3(CIC-3) on the growth and metastasis of cervical cancer cells

目的探讨氯离子通道蛋白-3(CIC-3)对宫颈癌细胞生长和转移的影响。方法通过免疫组织化学、qRT-PCR和Western blotting检测60例宫颈鳞癌组织和相应癌旁组织中CIC-3的表达。使用靶向CIC-3的siRNA转染SiHa细胞,并通过CCK-8、流式细胞术、伤口愈合实验和Transwell实验考察CIC-3对细胞生长和转移的影响。使用Western blotting检测细胞中PI3K、p-AKT、Bcl-2和p21的表达。此外,通过将转染CIC-3siRNA的SiHa细胞接种到BALB/c-nu/nu裸鼠背部复制体内肿瘤异种移植模型。结果免疫组织化学染色结果显示,CIC-3蛋白主要表达于子宫颈鳞状上皮细胞胞质区。癌组织CIC-3的阳性染色评分、CIC-3 mRNA相对表达量、CIC-3/β-actin相对表达量较癌旁组织高(P <0.05)。SiHa细胞CIC-3 mRNA相对表达量、CIC-3/β-actin相对表达量较H8细胞高(P <0.05)。不同肿瘤直径、TNM分期和是否有淋巴结转移患者CIC-3高表达率比较,差异有统计学意义(P <0.05)。下调CIC-3能抑制SiHa细胞的增殖、迁移和侵袭,并促进细胞凋亡(P <0.05)。在接种si-CIC-3转染的细胞后,裸鼠的肿瘤体积变小(P <0.05)。下调CIC-3能抑制PI3K、p-AKT和Bcl-2的蛋白表达,但促进p21的表达。结论 CIC-3在宫颈鳞癌中高表达,下调CIC-3可抑制宫颈鳞癌的生长和转移。CIC-3对宫颈鳞癌细胞凋亡的调控作用部分通过PI3K-AKT信号通路介导。

Objective To investigate the effects of chloride channel-3(CIC-3) on the growth and metastasis of cervical cancer cells. Methods The expression of CIC-3 in 60 cases of cervical squamous cell carcinoma tissues and adjacent tissues was detected by immunohistochemistry, quantitative real-time polymerase chain reaction(qRTPCR) and Western blotting. SiHa cells were transfected with small interfering RNA(siRNA) targeting CIC-3, and the effects of CIC-3 on cell growth and metastasis were investigated by cell counting kit-8, flow cytometry, wound healing assay, and transwell migration assay. Western blotting was used to detect the expression of phosphoinositide-3-kinase(PI3 K), p-AKT, B-cell lymphoma 2(Bcl-2) and p21. In addition, the in vivo xenograft tumor model was established by injecting SiHa cells transfected with CIC-3 siRNA into the backs of BALB/c-nu/nu nude mice.Results The CIC-3 was mainly expressed in cytoplasm of cervical squamous epithelial cells. The positive staining scores of CIC-3, relative expression of CIC-3 mRNA, and relative expression of CIC-3 to β-actin were higher in cervical squamous cell carcinoma tissues than those in adjacent tissues(P < 0.05). The relative expression of CIC-3 mRNA and that of CIC-3 to β-actin were higher in SiHa cells than in H8 cells(P < 0.05). The expression of CIC-3 differed in cervical squamous cell carcinoma tissues derived from patients with different tumor diameter, TNM stage and those with or without lymph node metastasis(P < 0.05). Down-regulation of CIC-3 inhibited the proliferation,migration, and invasion of SiHa cells, but promoted apoptosis(P < 0.05). After injection of cells transfected with CIC-3 siRNA, the tumor volume in nude mice was reduced(P < 0.05). Besides, down-regulation of CIC-3 inhibited the protein expression of PI3 K, p-AKT and Bcl-2, but promoted the expression of p21(P < 0.05). Conclusions CIC-3 is highly expressed in cervical squamous cell carcinoma. Down-regulating CIC-3 can inhibit the growth and metastasis of cervical squamous cell carcinoma. The regulation of CIC-3 on cell apoptosis in cervical squamous cell carcinoma is mediated in part by the PI3 K-AKT signaling pathway.