MiR-622 Regulates MAPK1 Signaling Pathway Effect On The Biological Function Of Ovarian Cancer Cells

Background and ObjectiveOvarian cancer is one of the three major tumors commonly seen in gynecology.Due to the initial symptoms of ovarian cancer patients and the lack of effective early diagnosis methods,the vast majority of patients are diagnosed at an advanced stage.Although the clinical skills of clinicians have improved,new equipment,and new drugs have been used in the treatment of ovarian tumors,the 5-year survival rate of advanced ovarian cancer is still around 25%.Therefore,it is imperative to study better early diagnosis and treatment of ovarian tumors.miRNA plays an important role in the regulation of cell proliferation,apoptosis,differentiation and pathogenesis.miRNA plays a dual biological function of "oncogene" and "tumor suppressor gene" in the process of tumor development and development.miR-622 can promote the proliferation of tumor cells such as liver cancer,gastric cancer and lung cancer,but its influence on the biological characteristics of ovarian cancer cells is still unclear.This study used mix in software to predict the target genes of miR-622 and explored miR-622 effects on the biological behavior of ovarian cancer cells.Materials and methodsqRT-PCR（reverse transcription polymerase chain reaction）was used to detect the expression of miR-622 in ovarian cancer cell lines SKOV3,OVCAR4,OVCAR8,TOV₂1G and normal ovarian epithelial cells IOSE80 and select the target cell line as the next experiment;The selected ovarian cancer cell lines were transfected with miR622-mi and mi-nc;miR622-in and in-nc,respectively,and the transfection efficiency was verified by qRT-PCR.The target genes that miR-622 may act on were predicted by the online websites such as Targetcar,miRDB and miRTarBase,and the expression of target genes in each group was verified by Western Blot.CCK8 detects the effect of miR-622 on the proliferation of ovarian cancer cells,cell flow assay to detect the effect of miR-622 on the apoptosis of ovarian cancer cells,Transwell and scratch test to detect the invasion and migration of ovarian cancer cells by miR-622 Results（1）The expression of miR-622 was up-regulated in ovarian cancer cell lines,from high to low,OVCAR4>SKOV3>OVCAR8>TOV-21 G.Compared with normal ovary cells IOSE80,the expression of miR-622 in ovarian cancer was statistically significant.Academic significance（P<0.05）.（2）OVCAR8 cells up-regulated by lipo3000（miR622-mi）and control group（mi-nc）and down-regulated group（miR622-in）and control group（in-nc）were collected and transfected into two groups.After RT-PCR,the results showed that the expression of miR-622 was significantly increased in the up-regulated group（miR622-mi group）,and the expression of miR-622 was significantly decreased in the down-regulated group（miR622-in group）,compared with the respective control groups.The difference was statistically significant（P<0.05）.（3）The binding region of MAPK1 and miR-622 was analyzed by biosignal.The results of Western Blot showed that the expression level of MAPK1 and ERK in the up-regulated group（miR622-mi）group was higher than that in the control group.（4）The results of CCK8 showed that there was no significant difference in the OD value between the up-regulated group（miR622-mi）and the control group（mi-nc）at 6h and 24h;the difference was statistically significant after 48h（P<0.05）;Compared with the control group（in-nc）,the OD value of the down-regulated group（miR622-in）was not statistically significant at 6h;the difference was statistically significant after 24h（P<0.05）.（5）Cell flow assay to detect apoptosis showed that there was no significant difference in apoptosis rate between the up-regulated group（miR622-mi）and the control group（mi-nc）;the down-regulated group（miR622-in）and Compared with the control group（in-nc）,the apoptosis rate was significantly higher than that of the control group.（6）The results of cell scratch test showed that the miR622-mi group had significantly higher cell migration ability than the control group at 24 h and 48 h after scratching compared with the control group（mi-nc）: down-regulation There was no significant difference in cell migration ability between the group（miR622-in）and the control group（in-nc）at 24 h and 48 h.（7）Transwell resuLts showed that there was no significant difference in cell invasion ability between the up-reguLated group（miR622-mi）compared with the control group（mi-nc）and the down-reguLated group（miR622-in）compared with the control group（in-nc）.Conclusions（1）The expression of microRNA622 was up-regulated in ovarian cancer cell lines.（2）MicroRNA622 promotes the proliferation and migration of ovarian cancer cells by up-regulating the MAPK1/ERK signaling pathway and inhibits the apoptosis of ovarian cancer cells.（3）MicroRNA622 had no effect on the invasive ability of OVCAR8 cells.