摘要
目的基于调控Janus蛋白酪氨酸激酶2/信号转导子和转录激活子3(JAK2/STAT3)通路探讨消痈乳康方对乳腺癌MCF-7细胞增殖、凋亡、侵袭的影响。方法实验分为空白对照组、消痈乳康方低浓度组、消痈乳康方中浓度组、消痈乳康方高浓度组,采用CCK8法检测细胞增殖情况,采用流式法检测细胞凋亡情况,采用插入式细胞穿透(Transwell)法检测细胞侵袭能力,采用RT-PCR技术检测细胞中JAK2/STAT3通路关键蛋白JAK2、p-JAK2、STAT3、p-STAT3 mRNA表达情况,采用Western blot法检测细胞中JAK2、p-JAK2、STAT3、p-STAT3蛋白表达情况。结果消痈乳康方各浓度组处理MCF-7细胞24 h、48 h、72 h后细胞抑制率和培养24 h后细胞凋亡率均明显高于空白对照组(P均<0.05),且消痈乳康方低、中、高浓度组细胞抑制率和细胞凋亡率逐渐增高,组间两两比较差异均有统计学意义(P均<0.05)。培养24 h后,消痈乳康方各浓度组细胞穿膜数均明显少于空白对照组(P均<0.05),且消痈乳康方低、中、高浓度组细胞穿膜数逐渐减少,组间两两比较差异均有统计学意义(P均<0.05)。培养48 h后,消痈乳康方低、中、高浓度组JAK2、p-JAK2、STAT3、p-STAT3 mRNA和蛋白相对表达量均明显低于空白对照组(P均<0.05),且消痈乳康方低、中、高浓度组JAK2、p-JAK2、STAT3、p-STAT3 mRNA和蛋白相对表达量逐渐降低,组间两两比较差异均有统计学意义(P均<0.05)。结论消痈乳康方能够有效抑制MCF-7细胞增殖、侵袭,促进细胞凋亡,其机制可能与抑制JAK2/STAT3信号通路激活有关。
Objective It is to investigate the effect of Xiaoyong Rukang Decoction(XYRKD)on the proliferation,apoptosis and invasion of breast cancer MCF-7 cells by regulating the Janus protein tyrosine kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3)pathway.Methods The experiment was divided into blank control group,low-concentration XYRKD group,medium-concentration XYRKD group and high-concentration XYRKD group.The cell proliferation was detected by CCK8 assay,the cell apoptosis was detected by by flow cytometry,the cell invasion ability was detected by Transwell method,the mRNA expression of JAK2/STAT3 pathway key proteins JAK2,p-JAK2,STAT3,p-STAT3 in cells was detected by RT-PCR technology,the expression of JAK2,p-JAK2,STAT3 and p-STAT3 proteins in cells was detected by Western blot method.Results The cell inhibition rate after MCF-7 treated for 24 h,48 h and 72 h and the cell apoptosis rate after cultured for 24 h in each concentration XYRKD group were significantly higher than those in the blank control group(all P<0.05),the cell inhibition rate and cell apoptosis rate increased gradually in the low,medium and high concentration groups,and there were significant differences compared with each other(all P<0.05).After 24 hours of culture,the quantity of cells penetrating the membrane in each concentration XYRKD group was significantly less than that in the blank control group(all P<0.05),and the quantity of cells penetrating the membrane in the low,medium and high concentration XYRKD groups gradually decreased,and there were significant differences compared with each other(all P<0.05).After 48 hours of culture,the relative expressions of JAK2,p-JAK2,STAT3,p-STAT3 mRNA and protein in the low,medium and high concentration XYRKD groups were significantly lower than those in the blank control group(all P<0.05),the relative expressions of JAK2,p-JAK2,STAT3,p-STAT3 mRNA and protein in the low,medium and high concentration XYRKD groups decreased gradually,and the differences between the two groups were statistically significant(all P<0.05).Conclusion XYRKD can effectively inhibit the proliferation,apoptosis and invasion of breast cancer MCF-7 cells.The specific mechanism of action may be related to its inhibition on JAK2/STAT3 signal pathway activation.
作者
唐众
凌洁
黄维芳
TANG Zhong;LING Jie;HUANG Weifang(The First Affiliated Hospital of Hunan University of Traditional Chinese Medicine,Changsha 410008,Hunan,China)
出处
《现代中西医结合杂志》
CAS
2022年第2期168-173,共6页
Modern Journal of Integrated Traditional Chinese and Western Medicine
基金
国家自然科学基金资助项目(81473680)
2020年度湖南省卫生健康委员会科研项目(202104011987)
湖南省教育厅科学研究项目(19C1395)。