摘要
目的 观察消岩汤含药血清对乳腺癌MCF-7细胞增殖、迁移和侵袭能力的影响,并探讨其机制是否与调控circRIP2和转化生长因子β(TGF-β)有关。方法 取对数生长期的MCF-7细胞,分为消岩汤含药血清4组、消岩汤含药血清2组、消岩汤含药血清1组、普通牛血清组、空白鼠血清组,分别加入浓度比为4∶2∶1的消岩汤含药血清、普通10%小牛血清及空白鼠血清100μL,孵育24 h。采用CCK-8法检测各组培养24、48 h的细胞增殖能力(以吸光度值表示),细胞划痕实验检测培养24、48、72 h的细胞迁移能力(以细胞迁移率表示),Transwell小室实验检测细胞侵袭能力(以穿膜细胞数表示),qRT-PCR法检测细胞circRIP2及RIP2、TGF-βmRNA表达,Western blotting法检测细胞丝氨酸苏氨酸激酶2(RIPK2)、TGF-β蛋白表达。结果 培养24 h,消岩汤含药血清4、2组细胞增殖能力均低于普通牛血清组(P均<0.05);培养48 h,消岩汤含药血清4、2、1组细胞增殖能力均低于普通牛血清组,消岩汤含药血清4组细胞增殖能力均低于空白鼠血清组(P均<0.05)。与普通牛血清组、空白鼠血清组同时间点比较,消岩汤含药血清4、2、1组24 h细胞迁移率及穿膜细胞数均降低,消岩汤含药血清4、2组48、72 h细胞迁移率均降低(P均<0.05)。消岩汤含药血清4组细胞circRIP2、TGF-βmRNA相对表达量均低于普通牛血清组、空白鼠血清组,RIP2mRNA相对表达量均低于普通牛血清组(P均<0.05)。消岩汤含药血清4组RIPK2蛋白相对表达量低于空白鼠血清组、消岩汤含药血清1组,TGF-β蛋白相对表达量低于普通牛血清组(P均<0.05)。结论 高浓度消岩汤含药血清可通过降低circRIP2、TGF-β表达而参与抑制乳腺癌MCF-7细胞的增殖、迁移和侵袭。
Objective To observe the effects of Xiaoyan decoction-containing serum on the proliferation,migration and invasion of MCF-7 cells,and to explore whether the mechanism is related to the regulation of circ RIP2 and transforming growth factor-β(TGF-β)or not.Methods MCF-7 cells in the logarithmic growth phase were divided into five groups:Xiaoyan decoction-containing serum group 4,Xiaoyan decoction-containing serum group 2,Xiaoyan decoctioncontaining serum group 1,bovine serum group,and mouse serum group.Xiaoyan decoction-containing serum(100μL)at a concentration ratio of 4∶2∶1,10%calf serum and mouse serum were added to the cells,then the cells were incubated for24 h.CCK-8 was used to detect the proliferation of MCF-7 at 24 and 48 h(OD value).The migration of MCF-7 at 24,48and 72 h(cell migration rate)was detected by Scratch test.Transwell test was applied to detect the invasion of cells at 24,48 and 72 h(number of trans-membrane cells).The m RNA level of circ RIP2,RIP2,and TGF-βwere detected by q RT-PCR.The protein levels of RIPK2 and TGF-βwere detected by Western blotting.Results After 24 h of culture,the cell proliferation abilities of Xiaoyan decoction-containing serum groups 4 and 2 were lower than that of the bovine serum group(both P<0.05).After 48 h of culture,the cell proliferation abilities of Xiaoyan decoction-containing serum groups4,2,and 1 were lower than that of the common bovine serum group(all P<0.05).The cell proliferation of Xiaoyan decoction-containing serum group 4 was lower than that of mouse serum group(P<0.05).Compared with the bovine serum group and mouse serum group at the same time point,the cell migration rate and the number of transmembrane cells decreased in the Xiaoyan decoction-containing serum groups 4,2 and 1 at 24 h(all P<0.05),and the cell migration rate decreased in the Xiaoyan decoction-containing serum groups 4 and 2 at 48 and 72 h(all P<0.05).The m RNA levels of circ RIP2 and TGF-βin Xiaoyan decoction-containing serum group 4 were lower than those in the bovine serum group and mouse serum group(all P<0.05),while the m RNA level of linear RIP2 in Xiaoyan decoction-containing serum group 4was lower than that in the bovine serum group(P<0.05).The protein level of RIPK2 in Xiaoyan decoction-containing serum group 4 was lower than those in mouse serum group and Xiaoyan decoction-containing serum group 1(P<0.05),and the protein level of TGF-βin Xiaoyan decoction containing serum group 4 was lower than that in the common bovine serum group(P<0.05).Conclusion High concentration of Xiaoyan decoction-containing serum inhibit the proliferation,migration and invasion of MCF-7 cells by reducing the expression of circ RIP2 and TGF-β.
作者
吕艳
耿强
韩燕燕
郑旭
李翀
兰福
赵杰
LYU Yan;GENG Qiang;HAN Yanyan;ZHENG Xu;LI Chong;LAN Fu;ZHAO Jie(Department of Pathology,The First Affiliated Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300381,China)
出处
《山东医药》
CAS
2022年第35期14-17,39,共5页
Shandong Medical Journal
基金
天津市教委科研计划项目(2017KJ160)。
