Abstract：Objective: To investigate the effects of gonadotropin inhibitory hormone (GnIH) on proliferation, apoptosis and PI3K/Akt/mTOR signaling pathway of human breast cancer MCF-7 cells based on GPR147 receptor. Methods: ① Human breast cancer cell line MCF-7 was cultured in vitro and divided into blank control group, control group and GnIH experimental group (0.1ng/mL, 1ng/mL, 10ng/mL, 100ng/mL, 1000ng/mL, 10000ng/mL). CCK-8 method was used to detect the cell proliferation; flow cytometry was used to detect the apoptosis rate; real time fluorescent quantitative PCR (QRT-PCR) and Western blot wereused to detect the effect of GnIH on apoptosis related proteins (Bcl-2, Bax, cytochrome C) at mRNA and protein levels . The expression of PI3K, Akt/p-Akt and mTOR proteins in Caspase-3, and p53 and PI3K/Akt/mTOR signaling pathways were detected. Western blot was used to detect the expression of GPR147 in MCF-7 cell line.② The cells were divided into PBS control group, DMSO control group, GnIH group and GnIH+RF9 inhibitor group for 24 hours. PBS 15 μL, DMSO 3μL, GnIH 10000 ng/mL and GnIH 10000 ng/mL+RF9 10μmol were added to the cells for 24 hours. The expression of Akt/p-Akt protein was detected by Western blot. Results: CCK-8 results showed that the GnIH experimental group 0.1ng/mL, 1ng/mL, 10ng/mL, 100ng/mL, 1000ng/mL cells compared with the control group, the difference was not statistically significant (P>0.05); Compared with the control group, GnIH10000ng/mL in the experimental group has a statistically significant difference (P<0.05). The apoptotic rates of control group, 1000ng/mL and 10000ng/mL were (7.76±1.57)% and (16.14±3.001)%, respectively, with significant difference (F=6.164, P=0.0351). The expressions of Bax, cytochrome c, caspase-3 and p53 were significantly up-regulated (P<0.05), while the expressions of Bcl-2 were significantly down regulated (P<0.05). In PI3K/Akt/mTOR signaling pathway, the protein expression of PI3K, Akt/pAkt and mTOR was significantly down regulated (P<0.01). GPR147 was found in MCF-7 cell line. When the drug concentration was 10000ng/mL, the receptor was activated compared with the control group (P<0.01). The expression of GPR147 in the experimental group was significantly higher than that in the control group (P<0.01). After the intervention, the expression of Akt/p-Akt protein was significantly higher in the group of GnIH +RF9 than in the group of GnIH (P<0.05). Conclusion: GnIH receptor GPR147 exists in breast cancer cell line MCF-7. GnIH may inhibit the proliferation and induce apoptosis of MCF-7 cells by binding to gpr147 receptor, and the occurrence of apoptosis may be related to the inhibition of PI3K/Akt/mTOR signaling pathway.
王凤霞, 魏萌, 司丽娜, 杨松鹤, 程露阳, 乔跃兵, 马秀艳. GnIH/RFRP-3通过受体GPR147抑制PI3K/AKT/mTOR通路诱导雌激素受体阳性人乳腺癌MCF-7细胞凋亡的研究[J]. 河北医学, 2021, 27(11): 1761-1766.
WANG Fengxia, WEI Meng, SI Lina, et al. GnIH/RFRP-3 Inhibits PI3K/Akt/MTOR Pathway Through GPR147 and Induces Apoptosis of Estrogen Receptor Positive Human Breast Cancer MCF-7 Cells. HeBei Med, 2021, 27(11): 1761-1766.
[1] Ullah MF.Breast Cancer:Current Perspectives on the Disease Status[J].Breast Cancer Metastasis and Drug Resistance,2019,1152:51~64. [2] 方玲,羊晓勤,李宏江.雌激素代谢与乳腺癌关系的研究进展[J].中国普外基础与临床杂志,2020,27(3):374~378. [3] Waks AG,Winer EP.Breast cancer treatment:a review[J].JAMA,2019,321(3):288~300. [4] 刘家硕,徐周,孔令泉,等.不同生理水平雌激素对激素受体阳性乳腺癌化疗敏感性的影响[J].第三军医大学学报,2020,42(15):1531~1535,1554. [5] Tian J M,Ran B,Zhang C L,et al.Estrogen and progesterone promote breast cancer cell proliferation by inducing cyclin G1 expression[J].Revista Brasileira De Pesquisas Med E Biol,2018,51(3):1~7. [6] 张鑫,霍孔林,宋星星,等.GnIH通过p38MAPK信号通路对猪卵巢颗粒细胞自噬与凋亡的影响[J].中国农业科学,2020,53(9):1904~1912. [7] Alzahrani AS.PI3K/Akt/mTOR inhibitors in cancer:at the bench and bedside[J].Seminars in Cancer Biology,2019,59:125~132. [8] Zhang W,Wang L,Yu X,et al.RFamide-related peptide-3 promotes alpha TC1 clone 6 cell survival likely via GPR147[J].Peptides,2018,107:39~44. [9] Zhang T,Han Z,Shi X,et al.Discovery of two novel branched peptidomimetics containing endomorphin-2 and RF9 pharmacophores:Synthesis and neuropharmacological evaluation[J].Bioorg Med Chem,2019,27(4):630~643. [10] 张延涛,孙勇,张远鹏.口虾蛄提取物对乳腺癌MCF-7细胞增殖及凋亡的影响及其可能机制[J].广西医学,2020,42(15):1995~2000. [11] Huang T C,Chiu P R,CHANG W T,et al.Epirubicin induces apoptosis in ost-eoblasts through death-receptor and mitochondrial pathways[J].Apoptosis,2018,23(3/4):226~236. [12] 郝剑.西黄丸通过ERα和HSP90在乳腺癌中发挥抗雌激素作用[D].天津市,天津医科大学,2018. [13] Yu Z,Jv Y,Cai L et al.Gambogic acid attenuates liver fibrosis by inhibiting the PI3K/AKT and MAPK signaling pathways via inhibiting HSP90[J].Toxicology and Applied Pharmacology,2019,371:63~73.
2021, Vol. 27 
冀公网安备13080202000677号