STOML-2过表达抑制人宫颈鳞癌Siha细胞凋亡

发布时间：2018-01-14 16:29

本文关键词：STOML-2过表达抑制人宫颈鳞癌Siha细胞凋亡　出处：《南方医科大学学报》2015年09期 　论文类型：期刊论文

【摘要】：目的研究STOML-2过表达抑制人宫颈鳞癌Siha细胞凋亡的机制,以期为宫颈癌的治疗开辟新的路径。方法用携带STOML-2基因的腺病毒感染Siha细胞作为过表达组,同时设立空白载体组及空白组作为对照,72 h后用Western blot检测STOML-2的过表达,并用MTT法检测STOML-2过表达对细胞增殖的影响。用IC50的顺铂处理各组细胞24 h后,利用荧光显微镜观察STOML-2过表达对细胞凋亡的影响,并利用流式细胞仪进一步分析细胞的凋亡率;采用Western blot检测线粒体凋亡途径相关蛋白caspase3、cleaved-caspase3、Bcl-2、Bax以及线粒体内外细胞色素C(Cyt C)表达量的变化。结果与空白组及空白载体组相比,Western blot结果显示过表达组STOML-2的表达量明显增加;MTT结果显示,STOML-2过表达后可明显促进Siha细胞的增殖。用IC50的顺铂处理细胞24 h后,荧光显微镜下,空白载体组凋亡细胞的细胞核大小不一,浓染致密的蓝色荧光显示核固缩,部分核裂解为凋亡小体,而过表达组细胞核多较完整,着色较浅,染色质密度均匀一致,并且流式细胞仪结果同样提示STOML-2过表达后细胞凋亡率明显降低。Western blot结果显示过表达组线粒体凋亡途径相关蛋白cleaved-caspase3、Bax以及线粒体外Cyt C表达水平降低,而caspase3、Bcl-2及线粒体内Cyt C的表达水平则相应升高。结论 STOML-2过表达可以促进Siha细胞增殖,其抑制凋亡的机制可能与线粒体凋亡途径相关。
[Abstract]:Objective to investigate the mechanism of overexpression of STOML-2 in inhibiting apoptosis of human cervical squamous cell carcinoma (Siha) cells. Methods Adenovirus-infected Siha cells carrying STOML-2 gene were used as overexpression group and blank vector group and blank group as control group. After 72 h, the overexpression of STOML-2 was detected by Western blot. The effect of STOML-2 overexpression on cell proliferation was detected by MTT assay. The cells in each group were treated with cisplatin of IC50 for 24 h. The effect of overexpression of STOML-2 on apoptosis was observed by fluorescence microscope, and the apoptosis rate was further analyzed by flow cytometry. The mitochondrial apoptosis-related protein caspase3, cleaved-caspase-3 and Bcl-2 were detected by Western blot. The expression of Bax and cytochrome C _ (Cyt C) in and out of mitochondria were compared with those in blank group and blank vector group. The results of Western blot showed that the expression of STOML-2 in overexpression group was significantly increased. The results of MTT showed that the over-expression of STOML-2 could significantly promote the proliferation of Siha cells. After treated with cisplatin of IC50 for 24 h, the cells were treated by fluorescence microscope. The nuclear size of apoptotic cells in blank vector group was different. The dense blue fluorescence showed nuclear pyknosis and partial nucleolysis into apoptotic bodies, while in overexpression group, the nuclei were more complete and light colored. Chromatin density is uniform. The results of flow cytometry also showed that the rate of apoptosis was significantly decreased after overexpression of STOML-2. The results of blot showed that the mitochondrial apoptosis pathway related protein cleaved-caspase3 in overexpression group. The expression of Bax and Cyt C in mitochondria decreased, while caspase3 decreased. The expression of Cyt C in Bcl-2 and mitochondria increased correspondingly. Conclusion the overexpression of STOML-2 can promote the proliferation of Siha cells. The mechanism of inhibiting apoptosis may be related to mitochondrial apoptosis pathway.
【作者单位】：南方医科大学南方医院;南方医科大学第三附属医院;新乡医学院第一附属医院;
【基金】：广东省自然科学基金(S2013010016194)
【分类号】：R737.33
【正文快照】： STOML-2是stomatin家族的新成员,具有stomatin家族共有的一致序列,但不具有N端跨膜区域,因此,STOML-2发挥功能的机制可能与其他成员不同[1-2]。STOML-2定位于线粒体内膜,其在食管癌、肺癌、子宫内膜癌等多种肿瘤均高表达[3-4],但其在宫颈癌组织中表达量的高低文献报道不一[5-9

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