靶向下调Cav3.1诱导人喉癌Hep-2细胞凋亡  

Cav3.1 targeted-knockdown induces apoptosis of human laryngeal squamous carcinoma Hep-2 cells

作　　者：江旋 许为青 潘春晨 Jiang Xuan;Xu Weiqing;Pan Chunchen(Dept of Otolaryngology-Head & Neck Surgery,The Affiliated Provincial Hospital of Anhui Medical University,Hefei 230001)

机构地区：[1]安徽医科大学附属省立医院耳鼻咽喉科,合肥230001

出　　处：《安徽医科大学学报》2021年第11期1723-1727,共5页Acta Universitatis Medicinalis Anhui

基　　金：“科大新医学”联合基金(编号:WK9110000053);安徽省自然科学基金(编号:1808085QH248);国家自然科学基金(编号:81800911)。

摘　　要：目的探讨Cav3.1在喉鳞癌组织和细胞中的表达以及靶向下调Cav3.1在喉鳞癌细胞增殖和凋亡的影响。方法应用免疫组织化学、RT-PCR和Western blot方法检测30例喉癌患者中Cav3.1在喉癌组织、正常切缘黏膜组织中的表达情况;siRNA瞬转技术构建靶向下调Cav3.1的Hep-2细胞系,应用Western blot检测各组细胞BAX、Cleaved-caspase-3、BCL-2的表达情况,应用CCK-8实验检测该喉癌细胞增殖的情况,Annexin V-PI染色技术检测喉癌Hep-2细胞凋亡的情况。结果相对癌旁组织,Cav3.1在喉癌组织中高表达(P<0.05);下调组Cav3.1在喉癌细胞中mRNA和蛋白的表达水平明显降低(P<0.05);下调Cav3.1可抑制喉癌细胞的增殖(P<0.05),促进喉癌细胞的凋亡(P<0.05),Western blot验证下调Cav3.1,BAX、Cleaved-caspase-3蛋白表达显著升高,BCL-2表达明显下降(P<0.05)。结论Cav3.1在喉癌组织和喉癌细胞系中高表达,下调Cav3.1抑制喉癌细胞的增殖,促进喉癌细胞凋亡。Objective To investigate the expression of Cav3.1 in laryngeal squamous cell carcinoma tissues and cells,and the effect of the proliferation and apoptosis in laryngeal squamous cell carcinoma by targeted-knockdown of Cav3.1.Methods Immunohistochemistry,RT-PCR and Western blot were used to detect the expression of Cav3.1 in laryngeal cancer tissues and normal mucosal tissues of incisional margin.Hep-2 cell lines of Cav3.1 targeted-knockdowned were constructed by siRNA transient technique.Western blot was used to detect the expression of BAX,Cleaved-caspase-3,BCL-2 in all groups.CCK-8 test was used to detect the proliferation of laryngeal carcinoma cell.Annexin V-PI staining was used to detect the apoptosis of laryngeal carcinoma cells in the targeted-knockdown group.Results Cav3.1 was highly expressed in 30 cases of laryngeal cancer compared to pericarcinomatous tissues(P<0.05).The expression of Cav3.1 mRNA and protein was decreased in si-Cav3.1 groups(P<0.05).In si-Cav3.1 group,the proliferation of laryngeal cancer cells decreased(P<0.05),the apoptosis of laryngeal carcinoma cells increased(P<0.05).The expression of BAX and Cleaved-caspase-3 was increased,and BCL-2 expression was decreased(P<0.05).Conclusion Cav3.1 is highly expressed in laryngeal cancer tissues and laryngeal cancer cell lines.Cav3.1 targeted-knockdown inhibits the proliferation and promotes apoptosis of laryngeal carcinoma cells.

关 键 词：喉鳞癌 HEP-2 Cav3.1 增殖 凋亡 

分 类 号：R739.65[医药卫生—肿瘤]