Lao Ling-Juan, Song Xin-Jiang, Xu Jia
Anorectal Surgery, Shaoxing Second Hospital, Shaoxing 312000, China.
Gastrointestinal Surgery, the First People Hospital of Yueyang, Yueyang 414000, China.
Zhongguo Zhong Yao Za Zhi. 2017 May;42(9):1730-1735. doi: 10.19540/j.cnki.cjcmm.20170222.014.
To analysis the relative expression of PTEN(phosphatase and tensin homology deleted on chromosome ten) in rectal cancer cell lines and the effects of resveratrol on cell proliferation and apoptosis of rectal cancer cells in vitro. Western blot was used to assess the protein expression of PTEN in rectal cancer cell lines and non-malignant colon epithelial cell line. After treating human rectal cancer cell line SW480 with different concentrations of resveratrol, the cellular proliferation was detected by cell counting Kit-8(CCK-8) assay. The apoptotic rate was analyzed by flow cytometry. The protein expressions of Caspase-3, PTEN, p53 and phosphorylase AKT(p-AKT) were detected by Western blot. shRNA(short hairpin RNA) was built to down-regulate PTEN expression in SW480 cells, and detect whether the own-regulated PTEN expression impact resveratrol's effect in growth inhibition and apoptosis induction of in vitro rectal cancer cells. Our results demonstrated that the relative level of PTEN was markedly decreased in Caco-2 and SW480, with statistically significant differences(P<0.001). Resveratrol inhibited the proliferation of rectal cancer cells markedly, and triggered apoptosis of SW480 cells. The protein expressions of Caspase-3, PTEN and p53 were all increased after being treated with resveratrol, while the expression of p-AKT was decreased after the treatment. The expression of endogenous PTEN was significantly decreased in shRNA PTEN-infected SW480 cells. Meanwhile, shRNA PTEN could obviously reduce SW480 cells' anti-proliferative and apoptotic effects. The experimental results suggested that resveratrol can be used as an anti-proliferative and apoptosis induction agent in rectal cancer through up-regulation of PTEN.
分析10号染色体缺失的磷酸酶和张力蛋白同源物（PTEN）在直肠癌细胞系中的相对表达以及白藜芦醇对直肠癌细胞体外增殖和凋亡的影响。采用蛋白质免疫印迹法检测PTEN在直肠癌细胞系和非恶性结肠上皮细胞系中的蛋白表达。用不同浓度的白藜芦醇处理人直肠癌细胞系SW480后，采用细胞计数试剂盒-8（CCK-8）法检测细胞增殖情况。通过流式细胞术分析凋亡率。采用蛋白质免疫印迹法检测半胱天冬酶-3、PTEN、p53和磷酸化蛋白激酶B（p-AKT）的蛋白表达。构建短发夹RNA（shRNA）下调SW480细胞中PTEN的表达，检测内源性PTEN表达的变化是否影响白藜芦醇对体外直肠癌细胞生长抑制和凋亡诱导的作用。结果显示，Caco-2和SW480细胞中PTEN的相对水平明显降低，差异具有统计学意义（P<0.001）。白藜芦醇显著抑制直肠癌细胞的增殖，并诱导SW480细胞凋亡。白藜芦醇处理后，半胱天冬酶-3、PTEN和p53的蛋白表达均增加，而p-AKT的表达降低。shRNA PTEN感染的SW480细胞中内源性PTEN的表达明显降低。同时，shRNA PTEN可明显减弱SW480细胞的抗增殖和凋亡作用。实验结果表明，白藜芦醇可通过上调PTEN作为直肠癌的抗增殖和凋亡诱导剂。
