摘要
目的:研究p-STAT3的活化在结肠癌中的表达与Snail、MMP2的相关性,及其在离体细胞实验中激活受抑后对结肠癌细胞迁移能力的影响并探讨其机制。方法:免疫组织化学染色检测p-STAT3与Snail、MMP2的表达及其相关性,分析三者与TNM分期、远处转移、淋巴结转移及分化程度之间的关系;MTT实验筛选AG490对增殖无影响的浓度和时间,并用该浓度和时间进行后续实验;采用Western blot法检测AG490抑制p-STAT3活化后STAT3、Snail、MMP2蛋白表达情况;划痕实验观察p-STAT3活化受抑后,结肠癌细胞迁移情况。结果:免疫组织化学染色结果表明,p-STAT3的表达与Snail、MMP2均存在相关性,且均与淋巴结转移相关(P<0.05)。在两个结肠癌细胞系中,通过MTT实验,选出10μM作为AG490的最佳作用浓度,并采用该浓度进行后续实验。AG490抑制p-STAT3活化后Snail、MMP2表达明显下降,而STAT3总蛋白表达无明显变化。且当AG490抑制p-STAT3活化后,细胞的迁移能力明显下降。结论:p-STAT3可以通过上皮间质转化(epithelial-mesenchymal transition,EMT)促进结肠癌的转移。
Objective： To determine p-STAT3 expression in 65 cases of colorectal cancer and its correlation with Snail and MMP2 and to explore the mechanism of p-STAT3 activation in colorectal cancer cells by inhibiting its activation in vitro. Methods： Immunohistochem- ical technique was adopted to examine the expression levels of p-STAT3, Snail, and MMP2. MTT assay was used to explore the effects of diluting AG490, which does not affect cell proliferation. Western blot was used to examine the expression levels of p-STAT3, STAT3, Snai1, and MMP2. Wound healing assay was employed to evaluate the migration of colorectal cancer cells. Results： p-STAT3 expression was significantly correlated with Snail and MMP2 expression. Moreover, p-STAT3 expression was significantly correlated with TNM stage, lymph node metastasis, and distant metastasis in colorectal cancer tissues （P〈0.05） but not with tumor differentiation. Treatment with 10 μM AG490 for 48 h did not significantly affect cell proliferation in the control and experimental groups （P〉0.05）. STAT3 expression was not significantly changed when p-STAT3 expression was inhibited by AG490. Meanwhile, the expression levels of Snail and MMP2 were down-regulated, and the migration ability of colorectal cancer cells was significantly reduced. Conclusion： p-STAT3 promotes colorectal cancer metastasis by regulating the process of epithelial-mesenchymal transition.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2017年第10期463-468,共6页
Chinese Journal of Clinical Oncology
基金
国家自然科学基金面上项目(编号:81572872)
国家自然科学基金重点项目(编号:81230050)资助~~
