摘要
目的探讨沉默PTEN基因后对结肠癌细胞增殖和侵袭影响的机理。方法 RT-PCR和Western blot法分析PTEN在4种结肠癌细胞株(HT-29、Wi Dr、Ca Co-2、Co Lo320细胞)中的表达,应用短基因干扰RNA(si RNA)技术合成PTEN si RNA并转染结肠癌细胞,Western blot法再次检测PTEN蛋白在结肠癌细胞株的表达。细胞增殖和侵袭实验分别检测PTEN基因沉默后对结肠癌细胞自身增殖和侵袭能力的影响。结果 PTEN在4种结肠癌细胞株中均有表达;4种结肠癌细胞转染PTEN si RNA后,Western blot法证实PTEN蛋白表达明显被抑制(P<0.01),结肠癌细胞的增殖和侵袭能力明显增强(P<0.01)。结论 PTEN基因的沉默能够明显增强结肠癌细胞的增殖和侵袭能力,增强PTEN基因可为临床靶向治疗结肠癌的转移提供新途径。
Objective To explore the influence mechanism of proliferation and invasion in colon cancer cell after silence ofphosphatase and tensin homolog deleted on chromosome 10 (PTEN) gene. Methods RT-PCR or Western blot method was used to detect the expression of PTEN mRNA or protein among four colon cancer cell lines (HT-29, WiDr, CaCo-2, and CoLo320 cell lines). Small interfering RNA (siRNA) was used to synthetize PTEN siRNA and transfect it into colon cancer cells. The expression of PTEN protein after transfecting was detected by Western blot. WST- 1 and invasion assay were used to examine the effects of PTEN siRNA silence on proliferation and invasion in colon cancer cells. Results PTEN mRNA and protein were expressed in all the four colon cancer cell lines. After PTEN siRNA transfected into the colon cancer cells, the expressions of PTEN proteins were inhibited in all the four colon cancer cell lines (P〈0.01), and the proliferation and invasion of colon cancer cells were enhanced significantly (P〈0.01). Conclusions PTEN siRNA play an important role in metastasis process of colon cancer via enhanced its proliferation and invasion. Therefore, the understanding biologic mechanisms for regulation of PTEN might enable better molecular target therapy of treating the colon cancer patients with metastasis.
出处
《中国普外基础与临床杂志》
CAS
2015年第5期544-548,共5页
Chinese Journal of Bases and Clinics In General Surgery
基金
国家自然科学基金项目(编号:81260325)
甘肃卫生行业科研计划项目(编号:GSWST2010-17)~~