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    竹节香附素A通过调节凋亡和EMT抑制口腔癌CAL27细胞生长和迁移

    Raddeanin A Inhibits Growth and Migration of Oral Cancer CAL27 Cells by Regulating Apoptosis and EMT

      摘要
    • 摘要:
      目的  研究竹节香附素A(raddeanin A，RA)通过调节凋亡和上皮细胞间充质转化(epithelial-mesenchymal transition，EMT)对口腔癌CAL27细胞生长和迁移的影响。
      方法 选取对数生长期的人口腔癌CAL27细胞，根据实验进行分组。采用CCK-8试验检测不同浓度RA对细胞增殖的影响；细胞克隆试验检测细胞增殖能力；Hoechst 33342染色试验和流式细胞术检测各组细胞凋亡情况；划痕试验检测各组细胞迁移能力的影响；Western blotting检测凋亡蛋白B淋巴细胞瘤-2(Bcl-2)、Bcl-2关联X蛋白(Bax)、活化半胱氨酸蛋白酶-3(Cleaved Caspase-3)及EMT通路相关蛋白钙黏蛋白E(E-cadherin)、钙黏蛋白N(N-cadherin)、波形蛋白(Vimentin)、锌指转录因子(Snail)表达。
      结果 CCK-8、克隆试验结果显示，与对照组相比，给药组细胞增殖能力呈浓度依赖性下降(P<0.05)；Hoechst 33342染色试验和流式细胞术结果显示，RA可以诱导CAL27细胞凋亡，抑制其生长(P<0.05)；划痕试验结果显示，细胞迁移能力降低(P<0.05)；Western blotting结果显示，凋亡蛋白Bcl-2表达降低，Bax、Cleaved Caspase-3表达显著增高(P<0.05)；EMT相关蛋白E-cadherin表达增高，N-cadherin、Vimentin、Snail表达显著降低(P<0.05)。
      结论 竹节香附素A可通过调节凋亡和EMT抑制口腔癌CAL27细胞生长和迁移。

       

      Abstract:
      OBJECTIVE  To investigate whether raddeanin A(RA) affects the growth and migration of oral cancer CAL27 cells through regulating apoptosis and epithelial-mesenchymal transition(EMT).
      METHODS  Human oral cancer CAL27 cells in logarithmic growth phase were selected and grouped according to the experiment. CCK-8 assay was used to detect the effects of RA on cell proliferation, cell cloning assay to detect cell proliferation ability, Hoechst 33342 staining assay and flow cytometry to detect apoptosis of cells, and scratch assay to detect the migration of cells. Western blotting was performed to detect the expression of apoptotic proteins B lymphoblastoma-2(Bcl-2), Bcl-2-associated X protein(Bax), Cleaved Caspase-3, and EMT-related proteins such as E-cadherin, N-cadherin, Vimentin and Snail.
      RESULTS  CCK-8 and clonogenic assay showed the cell proliferation of the administered group was decreased in a concentration-dependent manner(P<0.05 versus the control); Hoechst 33342 staining assay and flow cytometry showed that RA induced apoptosis and inhibited the growth of CAL27 cells(P<0.05 versus the control); and the scratch assay showed that the migration of the cells was decreased(P<0.05 versus the control). Western blotting showed that the expression of apoptotic protein Bcl-2 was significantly decreased, and the expression of Bax and Cleaved Caspase-3 significantly increased(P<0.05 versus the control). The expression of EMT-related protein E-cadherin was significantly increased, and the expression of N-cadherin, Vimentin and Snail significantly decreased(P<0.05 versus the control).
      CONCLUSION  Raddeanin A can inhibit growth and migration of oral cancer CAL27 cells through regulating apoptosis and EMT.

       

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