1.APOPTOSIS OF HUMAN OVARY GRANULOSA CELLS INDUCED BY THE METABOLITES ACYL-COA OF SATURATED FREE FATTY ACIDS
Yiming MU ; Hua ZHENG ; Baoa WANG
Medical Journal of Chinese People's Liberation Army 2001;0(08):-
To investigate the apoptotic effect of free fatty acids (FFAs) on human ovary granulosa cells, the granulosa cells were treated with FFAs in various concentrations for three days, and then the cell viability was determined by trypan blue exclusion method. DNA fragmentation was examined by DNA ladder formation and Annexin V GFP/PI staining of the cells. We also determined if the apoptotic effect of saturated FFAs was mediated by their acly CoA metabolites and measured the expression of apoptosis related genes, Bcl 2 and Bax by Western blot. The results indicated that saturated FFAs induced apoptosis of granulosa cell by their acly CoA metabolites in a dose dependent manner, and led to the down regulation of Bcl 2 and the up regulation of Bax. It is concluded that saturated FFAs can induce apoptosis of human ovary granulosa cells, suggesting that the sexual disorder of obese and insulin resistant women is related to it.
2.circNEIL3 regulates the proliferation, migration and invasion of breast cancer MDA-MB-231 cells through targeting miR-4784
ZHANG Baoa ; WANG Yanb ; REN Weimina
Chinese Journal of Cancer Biotherapy 2023;30(8):707-714
[摘 要] 目的:研究环状RNA nei样DNA糖化酶3(circNEIL3)和微小RNA(miR)-4784对乳腺癌细胞MDA-MB-231的增殖、迁移和侵袭的影响。方法:收集2018年1月至2019年12月在济南市中西医结合医院经组织病理诊断为乳腺癌并行手术切除的45例乳腺癌患者的癌组织和配对癌旁组织,qPCR法检测乳腺癌组织、癌旁组织中circNEIL3和miR-4784的相对水平。将circNEIL3的小干扰RNA(si-circNEIL3)、miR-4784模拟物、si-circNEIL3+miR-4784抑制物分别转染MDA-MB-231细胞,采用CCK-8法、平板克隆实验、划痕愈合实验、Transwell实验检测circNEIL3和miR-4784表达对细胞活力、克隆形成、迁移和侵袭的影响。双荧光素酶报告基因实验、RNA免疫沉淀(RIP)和RNA pull-down实验检测circNEIL3和miR-4784之间相互作用。结果:乳腺癌组织中circNEIL3呈高表达(P<0.05),miR-4784呈低表达(P<0.05)。干扰circNEIL3显著降低MDA-MB-231细胞活力、克隆形成数、划痕愈合率以及侵袭数(均P<0.05)。过表达miR-4784显著降低MDA-MB-231细胞活力、克隆形成数、划痕愈合率以及侵袭数(均P<0.05)。双荧光素酶报告基因实验、RIP和RNA pull-down实验均证实circNEIL3与miR-4784可直接结合,干扰circNEIL3能明显上调miR-4784表达(P<0.05),过表达circNEIL3能明显下调miR-4784表达(P<0.05)。抑制miR-4784表达部分逆转干扰circNEIL3对MDA-MB-231细胞活力、克隆形成、迁移和侵袭的抑制作用(P<0.05)。结论:干扰circNEIL3通过靶向上调miR-4784表达抑制MDA-MB-231细胞的增殖、迁移和侵袭。