摘要
目的:探究尿石素A(urolithinA,UA)对三阴性乳腺癌细胞恶性生物学行为的影响以及对蛋白激酶B(protein kinase B,AKT)/雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)信号通路的调控机制。方法:采用CCK-8法对人正常乳腺上皮细胞株MCF-10A和三阴性乳腺癌细胞MDA-MB-231探索UA的安全性和毒性;将人三阴性乳腺癌细胞MDA-MB-231随机分为MDA-MB-231组、MDA-MB-231细胞+尿石素A(UA)组、MDA-MB-231细胞+AKT激动剂(SC79)组、MDA-MB-231细胞+尿石素A+AKT激动剂(UA+SC79)组,采用平板克隆实验检测各组细胞的增殖活性;采用划痕实验和Tr-answell实验检测各组细胞的迁移和侵袭能力;采用流式细胞术检测各组细胞的凋亡情况;采用Western blot检测各组细胞中AKT/mTOR信号通路相关蛋白的活化水平。结果:确定7.5μmoL/L的UA既对正常乳腺细胞安全又对乳腺癌细胞有毒性,为本实验所用浓度。相比于MDA-MB-231组,UA组克隆形成数量(105.67±9.18)、细胞迁移率(12.92±2.13)、侵袭细胞数(56.33±3.68)以及细胞内p-AKT/AKT(0.12±0.02)和p-mTOR/mTOR值(0.16±0.03)均降低(P<0.05),而细胞凋亡率(22.1±1.75)升高(P<0.05);相比于MDA-MB-231组,SC79组克隆形成数量(222±17.57)、细胞迁移率(62.7±6.14)、侵袭细胞数(233.33±15.43)以及细胞内p-AKT/AKT(0.58±0.07)和p-mTOR/mTOR值(0.74±0.08)均升高(P<0.05),而细胞凋亡率(4.57±0.57)降低(P<0.05);SC79的加入逆转了UA对MDA-MB-231细胞恶性生物学行为的抑制作用(P<0.05)。结论:UA能够通过抑制AKT/mTOR信号通路抑制三阴性乳腺癌细胞的增殖、迁移、侵袭和抗凋亡等恶性生物学行为。
Objective:To investigate the effect of urolithin A(UA)on the malignant biological behavior of triple-negative breast cancer(TNBC)cells and its regulatory mechanism on the protein kinase B(AKT)/mammalian target of rapamycin(mTOR)signaling pathway.Methods:The safety and toxicity of UA were assessed on human normal breast epithelial cell line MCF-10A and TNBC cell line MDA-MB-231 using the CCK-8 assay.MDA-MB-231 cells were randomly divided into the following groups:MDA-MB-231,MDAMB-231+UA,MDA-MB-231+AKT activator(SC79),and MDA-MB-231+UA+AKT activator(UA+SC79).The proliferation activity of each group was detected by colony formation assay.Cell migration and invasion abilities were examined by wound healing and Transwell assays.Apoptosis was assessed by flow cytometry.The activation levels of AKT/mTOR signaling pathway-related proteins were detected by Western blot.Results:UA at a concentration of 7.5μmoL/L was determined to be safe for normal breast cells and toxic to breast cancer cells.Compared to the MDA-MB-231 group,the UA group showed decreased colony formation(105.67±9.18),cell migration rate(12.92±2.13),invasion cell count(56.33±3.68),and p-AKT/AKT(0.12±0.02)and p-mTOR/mTOR(0.16±0.03)levels(P<0.05),while the apoptosis rate(22.1±1.75)increased(P<0.05).In contrast,the SC79 group showed increased colony formation(222±17.57),cell migration rate(62.7±6.14),invasion cell count(233.33±15.43),and p-AKT/AKT(0.58±0.07)and pmTOR/mTOR(0.74±0.08)levels(P<0.05),and decreased apoptosis rate(4.57±0.57)(P<0.05).The addition of SC79 reversed the inhibitory effects of UA on the malignant biological behavior of MDA-MB-231 cells(P<0.05).Conclusion:UA can inhibit the proliferation,migration,invasion,and anti-apoptotic activities of TNBC cells by suppressing the AKT/mTOR signaling pathway.
作者
阿吉古
韩记真
陈艳艳
赖巧蓉
A Jigu;HAN Jizhen;CHEN Yanyan(Cancer Hospital,Xinjiang Medical University,Xinjiang Urumqi 830011,China)
出处
《河北医学》
CAS
2024年第8期1284-1290,共7页
Hebei Medicine
基金
新疆维吾尔自治区自然科学基金面上项目,(编号:2019D01C269)。
关键词
尿石素A
蛋白激酶B
雷帕霉素靶蛋白
三阴性乳腺癌
Urolithin A
Protein kinase B
Mammalian target of rapamycin
Triple-negative breast cancer
