摘要
目的基于EGFR-JNK通路,探讨RAA-11对人胃癌MGC-803细胞增殖和迁移的抑制作用。方法 MTT法检测RAA-11对人胃黏膜上皮GES-1细胞和人胃癌MGC-803细胞活力的影响;划痕实验检测RAA-11对人胃癌MGC-803细胞迁移能力的影响;实时荧光定量PCR检测RAA-11对人胃癌MGC-803细胞EGFR mRNA表达的影响;Western blot检测RAA-11对人胃癌MGC-803细胞凋亡相关蛋白caspase-3、Bcl-2、Bax及通路蛋白EGFR、JNK、p-JNK表达的影响。结果 MTT结果表明,与人胃黏膜上皮GES-1细胞相比,RAA-11明显抑制人胃癌MGC-803细胞的增殖(P<0.01),提示RAA-11对MGC-803细胞有选择作用;划痕实验结果提示,RAA-11能抑制人胃癌MGC-803细胞的迁移能力;实时荧光定量PCR结果提示,RAA-11降低了人胃癌MGC-803细胞EGFR mRNA的表达;Western blot结果提示,RAA-11上调人胃癌MGC-803细胞促凋亡相关蛋白caspase-3、Bax,并促进通路蛋白JNK、p-JNK的表达(P<0.01),下调抑凋亡相关蛋白Bcl-2的表达,并降低了通路蛋白EGFR的表达水平(P<0.01)。结论 RAA-11通过作用于EGFR-JNK通路,抑制人胃癌MGC-803细胞的增殖和迁移,并能够诱导其凋亡。
Aim To explore the effects of RAA-11 on the growth and migration of human gastric cancer cells MGC-803 via the EGFR-JNK pathway. Methods The effect of RAA-11 on the activity of human gastric mucosa cells GES-1 and human gastric cancer cells MGC-803 was observed by MTT assay. The effect of RAA-11 on the migration in human gastric cancer MGC-803 cells was detected by scratch test. The effect of RAA-11 on EGFR mRNA expression in human gastric cancer MGC-803 cells was detected by qRT-PCR. The changes of apoptosis-related proteins caspase-3, Bcl-2, Bax as well as pathway proteins EGFR, JNK, and p-JNK expression in human gastric cancer MGC-803 cells were assessed by Western blot. Results MTT results indicated that RAA-11 significantly inhibited the proliferation of MGC-803 cells in human gastric cancer compared with human gastric mucosa GES-1 cells(P<0.01), suggesting that RAA-11 had a selective effect on MGC-803 cells. The scratch result suggested that RAA-11 could suppress the migration of MGC-803 cells. The results of qRT-PCR indicated that RAA-11 decreased the expression level of EGFR mRNA in human gastric cancer MGC-803 cells. Western blot results suggested that RAA-11 up-regulated apoptosis-related proteins caspase-3 and Bax in human gastric cancer MGC-803 cells, promoted the expression of pathway proteins JNK and p-JNK( P < 0. 01),downregulated the expression of apoptosis-related proteins Bcl-2,and reduced the expression level of pathway protein EGFR( P < 0. 01). Conclusions RAA-11 can inhibit the proliferation and migration and induce apoptosis of human gastric cancer MGC-803 cells by the EGFR-JNK pathway.
引文
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