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

摘要:

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-3 siRNA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。

Abstract:

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 (qRT-PCR) 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 (PI3K), 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 PI3K, 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 PI3K-AKT signaling pathway.