菊藻丸含药血清调控PTEN对乳腺癌MCF-7细胞生长和转移的作用研究

Study on the effect of Juzao Pills drug-containing serum in regulating PTEN on the growth and metastasis breast cancer MCF-7 cells

目的 研究菊藻丸含药血清调控10号染色体上缺失的磷酸酶和张力蛋白同源物基因(PTEN)对人乳腺癌MCF-7细胞生长和转移的作用。方法 8周龄、体重(200±20)g的SPF级Wistar雌性大鼠12只,采用随机数字表法分为菊藻丸组和对照组,各6只。各组分别以5.4 g/(kg·d)菊藻丸和等量生理盐水灌胃,制备含药血清和空白对照血清。将培养好的人乳腺癌MCF-7细胞分为20%空白血清组、5%含药血清组、10%含药血清组、15%含药血清组和20%含药血清组,分别使用含20%空白对照血清和含5%、10%、15%、20%含药血清的培养基进行培养干预。采用CCK-8检测细胞增殖、集落形成实验观察克隆形成能力、TUNEL染色检测凋亡、流式细胞术检测周期分布、划痕实验和Transwell检测细胞迁移和侵袭能力,q PCR和Western blot检测PTEN m RNA和蛋白表达水平。结果 与20%空白血清组比较,10%、15%、20%含药血清组细胞增殖活性降低、克隆形成率减少、侵袭能力降低(P<0.05);5%、10%、15%、20%含药血清组细胞凋亡水平升高、迁移能力降低、细胞周期阻滞于G1期、PTEN的m RNA和蛋白表达水平均升高(P<0.05)。与5%含药血清组比较,10%、15%、20%含药血清组细胞增殖活性降低、克隆形成率降低、细胞凋亡水平升高、迁移能力降低、侵袭能力降低、细胞周期阻滞于G1期、PTEN m RNA和蛋白表达水平均升高(P<0.05)。结论 菊藻丸含药血清能抑制MCF-7细胞生长和转移,其机制可能与激活PTEN的表达有关。

Objective To study the effects of Juzao Pills drug-containing serum in regulating the expression of phosphatase and tensin homolog(PTEN)gene on the growth and metastasis of human breast cancer MCF-7 cells.Methods A total of 12 SPF female Wistar rats eight weeks old weighing(200±20)g were divided into Juzao Pills group and control group by random number table method,with six rats in each group.Each group was administered with 5.4 g/(kg·d)Juzao Pills or equal volume of saline by gavage to prepare drug-containing serum and blank control serum.The cultivated human breast cancer MCF-7 cells were divided into 20%blank control serum group,5%drug-containing serum group,10%drug-containing serum group,15%drug-containing serum group,and 20%drug-containing serum group,which were intervened with culture medium containing 20%blank control serum,5%,10%,15%,and 20%drug-containing serum,respectively.Then CCK-8 was used to detect cell proliferation,colony formation experiment was used to observe clonogenic ability,TUNEL staining was used to detect apoptosis,flow cytometry was used to detect cell cycle distribution,scratch test,and Transwell was used to detect cell migration and invasion abilities,qPCR and Western blot were used to detect PTEN mRNA and protein expression levels.Results Compared with 20%blank serum group,the cell proliferation activity,the cloning efficiency and the invasion ability were decreased in 10%,15%,and 20%drug-containing serum groups(P<0.05).In 5%,10%,15%,and 20%drug-containing serum groups,apoptosis level was increased,migration ability was decreased,cell cycle was blocked in G1 phase,mRNA and protein expression level of PTEN were increased(P<0.05).Compared with 5%drug-containing serum group,the cells in 10%,15%,and 20%drug-containing serum groups proliferation activity was decreased,cloning efficiency decreased,apoptosis level was increased,migration ability was decreased,invasion ability was decreased,cell cycle was blocked in G1 phase,and PTEN mRNA and protein expressionlevels increased(P<0.05).Conclusion Juzao Pills drug-containing serum could inhibit the growth and metastasis of MCF-7 cells,and its mechanism may be related to the activation of PTEN expression.