To Explore The Prognostic Factors And Mechanism Of Cervical Cancer Patients

Objective:1.To explore the independent risk factors of prognosis in patients with cervical cancer.To evaluate the prognostic value of LVSI in the stage IB-IIA cervical cancer and neoadjuvant chemotherapy in the narrow sense of local advanced cervical cancer.2.To search for the differential genes and related signal pathways that may affect the prognosis of cervical cancer,so as to provide new treatment strategies for drug-resistant cervical cancer patients.Methods:1.In this retrospective cohort study,patients with stage IB-IIA cervical cancer and local advanced cervical cancer in narrow sense at the Second Hospital of Jilin University from February 2014 to December 2016 were included.All patients with cervical cancer were finally treated with type C radical operation and pelvic lymphadenectomy.Hematoxylin eosin(H & E)staining was used to identify LVSI and evaluate the effect of LVSI on the prognosis of stage IB-IIA cervical cancer.Patients with locally advanced cervical cancer(?B2 and ?a2)in the narrow sense were grouped according to whether they received neoadjuvant chemotherapy(paclitaxel and platinum).The overall survival rate(OS)and progression free survival rate(PFS)were compared between the two groups.Kaplan–Meier curves were used to calculate the patient's survival.Survival was compared using the log-rank test,while risk factors for the prognosis were assessed by Cox regression analysis.2.The gene chip was used to screen the signal pathway related to the resistance of tumor to radiotherapy and chemotherapy.The target genes were identified by immunohistochemistry in patients with the same stage but different prognosis.The selected target gene was knocked out by lentivirus,the ability of proliferation and apoptosis of lentivirus transfected HeLa cells was detected by CCK-8 and flow cytometry with/without cisplatin combination,and the expression of P21,Bcl-2 and Bax were detected by Western blot.Results:1.The incidence of LVSI was positively associated with the depth of stromal invasion(P=0.009)and lymph node metastasis(LNM,P<0.001).LVSI is an independent factor that affects OS(P=0.009)and PFS(P=0.006)in patients with stage IB-IIA cervical cancer.The status of LNM is an independent factor that affects postoperative OS(P=0.005).2.Of the 144 patients of locally advanced cervical cancer,60 cases(41.7%)received neoadjuvant chemotherapy.84 cases(58.3%)did not receive neoadjuvant chemotherapy.The adverse reactions in the neoadjuvant chemotherapy group were mainly hematological toxic reactions,but tolerated.No grade ? ? adverse reactions were observed.Neoadjuvant chemotherapy did not significantly affect the progression-free survival(P=0.453)and overall survival(P=0.933)between the two groups.No factor was found to be independently associated with overall survival or progression-free survival(all P>0.05).3.Finally,HDAC6 and p21(CDKN1A)were screened out by gene chip.The results of immunohistochemistry showed that HDAC6 expressions were decreased,p21 expressions were increased and cell proliferation were inhibited in patients with good prognosis;HDAC6 expressions were increased and p21 expressions were decreased in patients with poor prognosis;The expressions of HDAC6 protein significantly reduced in HeLa cells transfected with HDAC6 lentivirus.Compared with the control group,the proliferation of HeLa cells transfected with HDAC6 lentivirus was significantly inhibited through CCK-8(P<0.05).The result showed that the apoptosis rate of HeLa cells transfected with HDAC6 lentivirus was significantly higher than that of the control group(P < 0.05)by Flow cytometry.The expressions of P21 in HeLa cells transfected with HDAC6 lentivirus was increased,the expressions of Bcl-2 were decreased,but the expressions of Bax were increased through Western blot.Compared with the control lentivirus with cisplatin group,HDAC6 lentivirus transfection combined with cisplatin reduced of the proliferation and significantly increased the apoptosis rate of HeLa cells,the difference was statistically significant.Conclusions:1.The incidence of lymphatic vessel infiltration is positively associated with the depth of stromal invasion and LNM.LVSI is an independent risk factor for the prognosis of stage IB-IIA cervical cancer.2.Compared with patients who underwent surgery with/without radiotherapy and/or chemotherapy,neoadjuvant chemotherapy using paclitaxel and the platinum does not improve the prognosis and lymph node metastasis rate of locally advanced cervical carcinoma.3.Transfection of HDAC6 knockdown virus significantly could reduce the expressions of HDAC6 protein in HeLa cells,which inhibiting the growth of cancer cells and inducing apoptosis of cancer cells.The combination of HDAC6 knockdown virus and chemotherapy drugs could inhibit the proliferation of HeLa cells and induce apoptosis.This may be related to the effect of HDAC6 knockdown in p21 proliferation and apoptosis signaling pathway.