| Ovarian cancer（OC）is the most deadly gynecological malignant tumor.Due to its hidden characteristics,many patients are already at an advanced stage when they are diagnosed after the appearance of clinical symptoms,which to a certain extent causes the overall survival rate of patients to be low.Therefore,it is very necessary to study the occurrence and development mechanism of ovarian cancer,find a new treatment directions for ovarian cancer patients,and provide a certain theoretical basis for clinical early diagnosis,effective treatment and prognosis evaluation of ovarian cancer.In recent years,studies have shown that compared with normal ovarian tissues,mi R-9 is significantly differentially expressed in ovarian cancer tissues,but the potential role of mi R-9 in the occurrence and development of ovarian cancer is still unclear.This study aims to explore whether the differential expression of mi R-9 in ovarian cancer is related to the tumor’s hypoxic microenvironment,and the specific underlying mechanism of mi R-9 in the occurrence and development of ovarian cancer Method（1）Collect clinical tissue samples: Collect ovarian cancer tissue and normal ovarian tissue from surgical patients from June 2020 to September 2021 in the Department of Obstetrics and Gynecology,Second Affiliated Hospital of Anhui Medical University,then use RT-q PCR experimental method to detect The expression level of mi R-9 in ovarian tissue.Western bloting and RT-q PCR were used to detect the expression of Hif-1α,PI3 K,P-PI3 K,AKT,P-AKT,m TOR,P-m TOR,GSK3β,and P-GSK-3β in ovarian tissues.（2）SKOV3 and HO-8910 cells were respectively cultured at 1% oxygen concentration for 0 H,0.5 h,1 h,2 h,4 h,8 h,16 h and 24 h,then the expression of mi R-9 was detected by RT-q PCR.The expression of Hif-1α,PI3 K,p-PI3 K,AKT,p-AKT,m TOR,p-m TOR,GSK3Β and p-GSK3β in different cell groups was detected by Western bloting and RT-q PCR.（3）SKOV3 and HO-8910 cells were cultured and divided into four groups.transfected with mi R-9 mimics,mi R-9 mimics NC,mi R-9 inhibitor,mi R-9inhibitor NC,respectively.Constructing mi R-9 mimics and inhibitor models and corresponding NC simulants.After the construction is successful,use Western bloting and RT-q PCR to detect the expression of Hif-1α,PI3 K,P-PI3 K,AKT,P-AKT,m TOR,P-m TOR,GSK-3β,P-GSK-3 in different groups of cells.（4）Treat SKOV3 and HO-8910 transfected with mi R-9 plasmid with LY294002,then use western bloting and RT-q PCR to detect the expression of Hif-1α,PI3 K,P-PI3 K,AKT,P-AKT,m TOR,P-m TOR,GSK-3β,P-GSK-3β in cells Results（1）Compared with normal ovarian tissue,the expression of mi R-9 in ovarian cancer tissue was significantly increased.Meanwhile,the expression of Hif-1α,P-PI3 K,P-AKT,P-m TOR and P-GSK-3 β were up-regulated in ovarian cancer tissues.（2）SKOV3 and HO-8910 cells were treated with Hypoxia,RT-q PCR results showed that the expression of mi R-9 was increased gradually and especially at 8H.at the same time,Hif-1α 、 P-PI3 K 、 P-AKT 、 Pm TOR and P-GSK3 β were also up-regulated in hypoxia group.（3）Western bloting and RT-q PCR showed that the expression of P-PI3 K,P-AKT,Pm TOR and P-GSK3 β in ovarian cancer cells SKOV3 and Ho-8910 transfected with mi R-9 mimics was up-regulated after over-expression mi R-9 and The expression of P-PI3 K,P-AKT,P-m TOR and P-GSK3 β was down-regulated in low-expression mi R-9.（4）Overexpressed mi R-9 can promote the expression of P-PI3 K,P-AKT,P-m TOR,and P-GSK3β.After LY294002 treated mi R-9 mimics-transfected ovarian cancer cells SKOV3 and HO-8910,these experiments showed that compared with the simple plasmid The group,the expression of P-PI3 K,P-AKT,P-m TOR,and P-GSK3β in the LY294002+mi R-9 mimics plasmid group was lower..ConclusionThe hypoxic microenvironment in tumor tissue can up-regulate the expression of mi R-9,and the over-expressed mi R-9 further promotes the occurrence and development of ovarian cancer by activating the PI3K/AKT/m TOR signaling pathway.Therefore,we speculate that down-regulation of mi R-9 may become a potential new treatment option for the treatment of ovarian cancer. |
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