1. Study of miR-18a/IGF-2/Akt pathway regulating the proliferation, migration and invasion of ovarian cancer cells
Guancui LAN ; Yang GAO ; Yongju YE
Chinese Journal of Endocrine Surgery 2019;13(6):505-509
Objective:
To study the role of miR-18a in regulating the proliferation, migration and invasion of ovarian cancer cells through IGF-2/Akt pathway.
Methods:
Ovarian cancer SKOV3 cells were cultured and grouped. The negative control group were not transfected with the plasmid, the blank plasmid group were transfected with the blank pcDNA3.1 plasmid, the IGF-2 plasmid group was transfected with the pcDNA3.1 plasmid expressing IGF-2, and the IGF-2 plasmid+L294002 group was transfected with pcDNA3.1 plasmid expressing IGF-2 and treated with Akt inhibitor LY294002. The upstream miRNA of IGF-2 was analyzed by bioinformatics method and verified by the double luciferase reporter gene method. The effect of overexpressing miR-18a on the expression of IGF-2 was observed.
Results:
Transfection of IGF-2 expression plasmid can increase the expression level of IGF-2 in cells and the content of IGF-2 in culture medium (
2.Exploring the effect of Xinjia Erjia Dihuang Tang on follicular development in PCOS model rats based on the Kisspeptin/GPR54 system
Mingqing SHI ; Jin WANG ; Xiaoyu XU ; Guancui LAN
China Modern Doctor 2024;62(18):90-95
Objective To investigate the development of follicles in polycystic ovary syndrome(PCOS)model rats treated with Xinjia Erjia Dihuang Tang and its possible mechanism.Methods Twenty-eight female SD rats with regular estrous cycle were selected and randomly divided into normal control group,model group,Chinese medicine group and Western medicine group,with 7 rats in each group.In addition to the normal control group,the other three groups were continuously given 0.1mg/(kg·d)of letrozole-carboxymethyl cellulose suspension to construct the PCOS rat model.Since the 22nd day,the traditional Chinese medicine group was given 5.268g/(kg·d)of Xinjia Erjia Dihuang Tang,the western medicine group was given 0.286mg/(kg·d)of ethinylestradiol cyproterone tablet,and the normal control group and model group were given 10ml/(kg·d)of distilled water.After 3 weeks,the ovarian coefficients of the rats in all groups were compared,and the morphological changes of the ovarian tissues of the rats in all groups were observed by hematoxylin-eosin staining.The serum hormones of the rats in all groups were detected by enzyme-linked immunosorbent assay,and the ovarian kisspeptin(Kp)and G-protein-coupled receptors(GPR54)protein expression levels were detected by Western blot.Results Compared with the normal control group,the follicles in the ovary of the model group showed more cystic dilatation and atresia,and the number of granular cell layers decreased,the ovarian coefficient of the model group was increased;Serum levels of testosterone(T),luteinizing hormone(LH)and Kp were increased in model group;Serum levels of follicle stimulating hormone(FSH)and estradiol(E2)and protein expressions of Kp and GPR54 in ovarian tissue of rats in the model group were significantly decreased(P<0.05).After treatment with Chinese medicine,compared with the model group,the number of follicles in the Chinese medicine group decreased,the layer number of granulosa cells increased,there were nearly mature follicles,and a small amount of corpus luteum existed,serum LH,T and Kp levels were decreased,serum FSH and E2 levels and ovarian Kp and GPR54 protein expressions were significantly increased(P<0.05).Conclusion The development of follicles in PCOS model rats was improved by the intervention of Xinjia Erjia Dihuang Tang.The therapeutic mechanism may be to affect the release of FSH and LH by regulating Kisspeptin/GPR54 system,so as to affect hormone content,adjust ovarian function,and improve follicle development and ovulation ability.