周晓谦 金星姬 杨竹生 杨秀敏

[摘要]目的：探讨百里香醌对人皮肤鳞状细胞癌A431细胞增殖、凋亡的影响及可能机制。方法：将A431细胞分为对照组，低、中、高浓度实验组，对照组采用DMEM培养，而低、中、高浓度实验组分别采用含1、15、30μg/ml百里香醌的DMEM培养。MTT法检A431细胞增殖;台盼蓝法观察A431细胞存活;划痕试验检测各组A431细胞迁移;免疫荧光法检测Bax和Bcl-2蛋白表达;Westernblot检测各组A431细胞中Bax、Bcl-2、Caspase-3、Cyto C和PARP蛋白表达。结果：百里香醌能有效抑制人皮肤鳞状细胞癌A431细胞增殖，且呈浓度及时间依赖性;台盼蓝法显示，百里香醌浓度增加使A431細胞死亡率上升，呈浓度依赖性;与对照组比较，低、中、高浓度实验组A431细胞迁移能力降低;与对照组比较，经百里香醌处理后A431细胞Bax表达增加，Bcl-2表达降低;Western blot检测结果显示，与对照组比较，不同浓度百里香醌处理后A431细胞Bax、cyto-C、caspase-3和PARP表达水平增加，而Bcl-2表达下降。结论：百里香醌具有抑制A431细胞增殖、侵袭以及诱导凋亡的作用，其机制可能是Bax/Bcl-2比值升高，促使释放至胞浆中Cyto C增加，进而激活Caspase-3以及PARP蛋白有关。

[关键词]百里香醌;人皮肤鳞状细胞癌;增殖;凋亡;迁移

[中图分类号]R739.5    [文献标志码]A    [文章编号]1008-6455（2021）06-0114-04

Thymoquinone Regulates Caspase-3/Bax/Bcl-2 Signaling Pathway and Induces Proliferation and Apoptosis of Cutaneous Squamous Cell Carcinoma

ZHOU Xiao-qian，JIN Xing-ji，YANG Zhu-sheng，YANG Xiu-min

（Department of Dermatology，Beijing Tongren Hospital，Capital Medical University，Beijing 100730，China）

Abstract： Objective  To investigate the effect of thymoquinone on the proliferation and apoptosis of human cutaneous squamous cell carcinoma A431 cells and its possible mechanism. Methods  A431 cells were divided into the control group and the low， medium and high concentration experimental groups. The control group was cultured with DMEM， while the experimental-L， M， H groups were cultured with DMEM containing 1， 15 and 30 μg/ml thymoquinone， respectively. The proliferation of A431 cells was detected by MTT. The survival of A431 cells was observed by trypan blue method. The migration of A431 cells was detected by scratch test. The expression of Bax and Bcl-2 protein was detected by immunofluorescence. The expression of Bax， Bcl-2， Caspase-3， Cyto C and PARP protein in A431 cells were detected by Western blot. Results  Thymoquinone can effectively inhibit the proliferation of human skin squamous cell carcinoma A431 cells in a concentration and time-dependent manner. Trypan blue assay showed that the mortality of A431 cells increased with the increase of thymoquinone concentration. Compared with the control group， the migration ability of A431 cells in the experimental-L， M， H groups was decreased. Compared with the control group， the expression of Bax increased and the expression of Bcl-2 decreased in A431 cells treated with thymoquinone. Compared with the control group， the expression levels of Bax， Cyto-c， Caspase-3 and PARP were increased in A431 cells treated with different concentrations of thymoquinone， while the expression of Bcl-2 was decreased. Conclusion  Thymoquinone can inhibit the proliferation， invasion and induce apoptosis of A431 cells. The mechanism may be related to the increase of the ratio of Bax/Bcl-2， the increase of Cyto C released into the cytoplasm， and the activation of Caspase-3 and PARP protein.

Key words： thymoquinone; human cutaneous squamous cell carcinoma; proliferation; apoptosis; migration

人体皮肤是最直接与外界接触的器官，有助于吸收、分泌和免疫，是人体抵御外界有害物质侵入的第一道防线[1]。近年来，皮肤鳞状细胞癌（Cutaneous squamous cell carcinoma，cSCC）在皮肤非黑色素细胞性恶性肿瘤中发病率居第二位，且呈增高趋势[2]。然而，许多公众对皮肤癌缺乏认识，导致延误诊断，影响及时有效的治疗，严重影响患者的健康和生活。……