中国麻风皮肤病杂志 ›› 2021, Vol. 37 ›› Issue (8): 499-505.doi: 10.12144/zgmfskin202108499

• 论著 • 上一篇    下一篇

抗增殖蛋白2在皮肤鳞状细胞癌中的表达及其对A431细胞增殖的影响

孔晓芸,缪旭,张冬梅,崔恒祥,施健,刘念念   

  1. 南通大学第二附属医院,江苏南通,226000
  • 出版日期:2021-08-15 发布日期:2021-07-20
  • 通讯作者: 缪旭,E-mail: mx730129@163.com

Expression of prohibitins 2 in cutaneous squamous cell carcinoma and its effect on proliferation of cutaneous squamous cell carcinoma cell line A431

KONG Xiaoyun, MIAO Xu, ZHANG Dongmei, CUI Hengxiang, SHI Jian, LIU Niannian   

  1. Second Affiliated Hospital of Nantong University, Nantong 226000, China
  • Online:2021-08-15 Published:2021-07-20
  • Contact: MIAO Xu, E-mail: mx730129@163.com

摘要: 目的:观察抗增殖蛋白2在皮肤鳞状细胞癌、鲍温病、日光性角化病组织中的表达情况及其对A431细胞增殖作用的影响。方法:采用免疫组织化学方法检测抗增殖蛋白2在40例皮肤鳞状细胞癌、18例鲍温病、17例日光性角化病组织以及9例正常皮肤组织中的表达水平。采用CRISPR-Cas9法构建两组靶向敲除抗增殖蛋白2的皮肤鳞状细胞癌A431细胞模型(KO299组及KO320组),使用Western印迹法证实其敲除效果,通过CCK8法及细胞克隆形成实验分析其对细胞增殖的影响。使用Western印迹法对AKT蛋白表达及其ser473位点磷酸化产物水平进行分析。 结果:抗增殖蛋白2在皮肤鳞状细胞癌(90.00%)、鲍温病(66.70%)、日光性角化病(41.18%)的表达高于正常组织(0.00%)(均P<0.05),皮肤鳞状细胞癌抗增殖蛋白2表达高于日光性角化病(P<0.05)。在不同性别、年龄、部位、病程、肿瘤直径、浸润深度、分化、Broder分级的皮肤鳞状细胞癌组织中抗增殖蛋白2的表达差异无统计学意义(均P>0.05)。两组抗增殖蛋白2敲除组A431细胞的蛋白表达水平、细胞活力、克隆形成数显著低于对照组 (均P<0.05)。p-AKT表达随抗增殖蛋白2表达水平的降低而显著下调(均P<0.01)。结论:抗增殖蛋白2的表达可能促进了皮肤鳞状细胞癌的发生,并且其机制可能与促癌基因AKT蛋白活化相关。

关键词: 抗增殖蛋白2, 皮肤鳞状细胞癌, 细胞增殖, 蛋白激酶B

Abstract: Objective: To detect the expression of prohibitins 2 in cutaneous squamous cell carcinoma, Bowen's disease and actinic keratosis, and to evaluate the effect of prohibitins 2 knockout on the proliferation of cutaneous squamous cell carcinoma cell line A431. Methods: The expression of prohibitins 2 in 40 cases of cutaneous squamous cell carcinoma, 18 cases of Bowen's disease, 17 cases of actinic keratosis and 9 cases of normal skin tissue were detected by immunohistochemistry. Prohibitins 2 knockout A431 cells were generated by CRISPR/Cas9 technology and the knockout effect was confirmed by Western blot. The effect on cell proliferation was measured by CCK8 and cell clone formation experiment. Additionally, the phosphorylation level of AKT at Ser473 was semiquantitatively analyzed by Western blot. Results: The nuclear expression of prohibitins 2 in cutaneous squamous cell carcinoma (90.00%), Bowen's disease (66.70%) and actinic keratosis (41.18%) was higher than that in normal skin tissue (0.00%)(Ps<0.05), and the expression of prohibitins 2 in cutaneous squamous cell carcinoma was higher than that in actinic keratosis (P<0.05).The expression rates of prohibitins 2 in cutaneous squamous cell carcinoma, Bowen's disease, actinic keratosis and normal skin tissue were decreased gradually and there was a significant difference among these 4 groups (P<0.01). There was no significant difference of the expression of prohibitins 2 in different sex, age, location, course of disease, tumor diameter, depth of invasion, differentiation, Broder grade (Ps>0.05). The expression of prohibitins 2, the cell viability, the numbers of clone formation in two prohibitins 2 knockout groups  were lower than that in control group, with significant differences (Ps<0.05). The expression of p-AKT was significantly down-regulated with the decrease of prohibitins 2 expression level (P<0.05). Conclusion: The expression of prohibitins 2 may promote the occurrence of cutaneous squamous cell carcinoma, and its mechanism may be related to the activation of oncogene AKT.

Key words: prohibitins 2, cutaneous squamous cell carcinoma, cell proliferation, AKT