| Purpose:Platinum-based combined chemotherapy played important role in the treatment of ovarian cancer.Platinum resistance is the main factor that leads to poor prognosis of ovarian cancer.So it is of great significance to find molecular targets for predicting and reversing platinum resistance.Previously we found that a novel E3 ubiquitin ligase HECTD3 is a regulator of cell apoptosis wh can protect cancer cell survival and confer cancer cell resistance to apoptosis.Besiedes,HECTD3 could regulate platinum sensitivity in breast cancer and cervival cancer in vitro.In this paper,the expression of HECTD3 in ovarian cancer and the underlying regulation mechanisms of platinum resistance were studied.Methods:The study is divided into four parts.1)150 patients of advanced serous ovarian adenocarcinoma that collected from May 2007 to January 2013 in our hospital were included in the retrospective study,and their surgical specimens of ovarian tumor were made into tissue microarray.Expression of HECTD3 was evaluated via immunohistochemical staining,and was compared with clinical-pathologic characteristics.Multivariate analysis was used to evaluate prognostic factors,and PFS and OS were anaylized by Kaplan-Meier method.2)The difference of HECTD3 expression between SKOV3 and cisplatin-induced drug-resistant cell line SKOV3R(DDP)was compared by western blotting(WB).Besides,the differences in apoptosis induced by carboplatin between SKOV3 and SKOV3R were detected by flow cytometry.The virus containing the HECTD3 knockdown vector and the control vector was used to infect SKOV3R cells respectively.After screening and constructing the stable cell line,the knockdown level of HECTD3 was confirmed by WB.Then,the difference of carboplatin-induced apoptosis was detected by flow cytometry using Annexin V/PI staining method.The bilateral axillary subcutaneous tumo rmodel of mice was constructed by HECTD3 knockdown and control SKOV3R cell line.Carboplatin was given to compare the effect of tumor inhibition.3)The expression of HECTD3 and pSTAT3 was detected by WB in SKOV3 and A2780 cell lines treated with Her2 inhibitors(Lapatininb,Neratinib and Herceptin).Moreover,SKOV3 and A2780 cells were transfected with siRNA,in order to transiently knock down Her2,then,expression of Her2,pSTAT3 and HECTD3were evaluated by western blot.Then,STAT3 knocking-down siRNA and STAT3overexpressing virus were used to infect SKOV3 cells respectively.WB was used to detect the expression of pSTAT3 and HECTD3.Correlation analysis of HECTD3 and Her2 expression was analyzed via IHC of tissue microarray.In addition,STAT3 was used to locate the sequence of HECTD3 promoter activity by double luciferase reporter assay.4)In vitro,apoptosis differences induced by carboplatin,lapatinib or combined application respectively in SKOV3 cell lines were dectected by flow cytometry using Annexin V/PI staining method.While in vivo,SKOV3 cells were used to construct the subcutaneous tumor model of mice,and the oboved metioned drug applicaiton were performed on sperated groups.Finally,the therapeutic effect was compared by tumor weight.Results:In this study,we found that platinum-sensitive or resistant relapse is closely related with expression of HECTD3 through IHC and clinical and pathological data analysis(P<0.01).The higher HECTD3 expression in the ovarian serous cancer,the poorer prognosis is.The expression of HECTD3(P<0.05),FIGO staging(P<0.05)and residual tumor size(P<0.05)were the independent prognostic factors of PFS.While the expression of HECTD3 alone(P<0.05)was independent prognostic factor of OS.2)SKOV3R exhibits higher HECTD3 expression than its parental cell line SKOV3R.SKOV3R was more resistant to carboplatin than SKOV3 in vitro.HECTD3 depletion promotes carboplatin-induced apoptosis both in ovarian cancer cell model in vitro and also in mice model in vivo.3)HECTD3 was found positively correlated with the expression of Her2 in ovarian cancer tissue(spearman correlation coefficient is 0.214,P=0.007).Furthermore,Her2 can regulate HECTD3 expression through activating STAT3.Her2 inhibitor,such as Lapatinib etc,could down-regulate HECTD3 expression at certain condition thus promoting chemosensitivity of ovarian cancer cells to carboplatin.4)Lapatinib combinated with carboplatin significantly inhibited serous ovarian carcinoma tumor growth compared with either drug alone in vitro(A2780:CBP-H P=0.047;SKOV3:CBP-L P=0.045,CBP-H P=0.032)and in vivo(CBP+L vs CBP P<0.0001,CBP+L vs L P=0.0001).Conclusions:In conclusion,HECTD3 is positively correlated with platinum resistance and is regarded as an independent prognotstic factor of serous ovarian adenocarcinoma.Down-regulating HECTD3 expression could increase the sensitivity of ovarian cancer to platinum drugs.In addition,HECTD3 was positively regulated through STAT3 in Her2 signal pathway and was positively correlated with Her2 at the tissue level.Thus,Her2 inhibitors can be used to sensitize platinum-based chemotherapy.Taken together,HECTD3 has great potential becoming a molecular predictor and therapeutic target for serous ovarian adenocarcinoma.Objectives:The study aimed to evaluate securiry and clinical prognosis of nerve-sparing radical hysterectomy(NSRH)for locally advanced cervival cancer(LACC).Methods:A retrospective cohort analysis was conducted in the department of gynecological oncology,National Cancer Center/Cancer Hospital,Chinese Academy of Medical Sciences.Patients with locoally advanced squamous cervical cancer(stage in Ib2 and IIa2)were recruited between January 1,2008 and December 31,2014.Of all the patients,69 cases underwent NSRH,while other 320 cases underwent conventiaon radical hysterectomy(RH).We compared patients’ general characteristics and surgical parameters between two groups.After median 67 months’ follow-up,progression-free and overall survival as well as recovery of bladder function in recent and future was evaluated.Results:Basic clinical and pathological information on two groups were matched.Multivariate analysis showed that pelvic lymphatic metastasis was the independent prognostic factor of DFS(HR=1.31,95%CI 为 1.01～1.68,P=0.040)as well as OS(HR=1.30,95%CI为 1.01～1.67,P=0.046)for locally advanced cervival cancer.Preserving pelvic splanchnic nerves or not has no effect on the PFS(P=0.746)nor OS(P=0.910)for locally advanced cervical cancer.Compared with patients treated with RH,patients who underwent NSRH presented longer operation time(262±51min vs.245±52min,P<0.01),but less blood loss(366±302ml vs.457±301ml,P<0.05),shorter urethral catheterization time(11.0±8.9 days vs.15.5±6.6 days,P<0.01)and lower proportion of bladder dysfunction 1 year after surgery(6.1%vs.16.3%,P<0.05).Conclusions:NSRH is a safe and feasible surgical procedure for patients with locally advanced cervical cancer.More importantly,compared with RH,NSRH not only demestrateds similar prognosis,but also impove surgical quality as well as bladder function for patients with LACC. |
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