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目的研究组蛋白去乙酰化酶6(histonedeacetylase6,HDAC6)siRNA对皮肤鳞状细胞癌(简称鳞癌)SCL-1细胞增殖和凋亡的影响及分子机制。方法将SCL-1细胞分为三组:A为未处理组;B为siRNA对照组;C为HDAC6siRNA转染组。利用脂质体2000分别介导HDAC6siRNA和对照siRNA至SCL-1细胞中,利用Westernblot研究HDAC6siRNA对SCL-1细胞中HDAC6蛋白表达的影响。采用CCK-8试剂和流式细胞术分别研究HDAC6siRNA对SCL-1细胞增殖和细胞凋亡的影响,进一步利用WesternBlot技术分析细胞增殖相关蛋白P21和细胞凋亡相关蛋白Bcl-2及Bax的表达。结果 HDAC6siRNA转染组中HDAC6蛋白表达显著低于未处理组和siRNA对照组,差异具有统计学意义(P<0.05)。与未处理组和siRNA对照组相比,HDAC6siRNA转染组中SCL-1细胞的增殖明显受到抑制(P<0.05)。HDAC6siRNA转染组中SCL-1细胞的早期凋亡率明显高于未处理组和siRNA对照组,差异具有统计学意义(P<0.05)。HDAC6siRNA转染组中Bcl-2表达明显下调,而P21和Bax表达显著上升。结论 HDAC6可能在皮肤鳞癌的发生发展中发挥重要作用,下调皮肤鳞癌中HDAC6的表达有望成为有用的分子治疗靶点。
Objective To investigate the effects of histone deacetylase 6 (siRNA) on the proliferation and apoptosis of SCL-1 cells and its molecular mechanism in cutaneous squamous cell carcinoma (SCC). Methods SCL-1 cells were divided into three groups: A was untreated group; B was siRNA control group; C was HDAC6 siRNA transfection group. HDAC6 siRNA and control siRNA were respectively introduced into SCL-1 cells by lipofectamine 2000. The effect of HDAC6 siRNA on HDAC6 protein expression in SCL-1 cells was analyzed by Western blot. The effects of HDAC6 siRNA on proliferation and apoptosis of SCL-1 cells were studied by CCK-8 reagent and flow cytometry respectively. The expression of cell proliferation-related protein P21 and apoptosis-related proteins Bcl-2 and Bax were further analyzed by Western Blot. Results The expression of HDAC6 protein in HDAC6 siRNA transfection group was significantly lower than that in untreated group and siRNA control group (P <0.05). The proliferation of SCL-1 cells in HDAC6 siRNA transfection group was significantly inhibited compared with untreated and siRNA control groups (P <0.05). The early apoptosis rate of SCL-1 cells in HDAC6siRNA transfection group was significantly higher than that in untreated group and siRNA control group, the difference was statistically significant (P <0.05). The expression of Bcl-2 in HDAC6siRNA transfection group was significantly down-regulated while the expression of P21 and Bax was significantly up-regulated. Conclusion HDAC6 may play an important role in the development of squamous cell carcinoma. Down-regulation of HDAC6 expression in squamous cell carcinoma of the lung is expected to be a useful target for molecular therapy.