IL-1β inhibitor sensitizes to olaparib in homologous recombination deficiency proficient ovarian cancer cells / 中华妇产科杂志

Objective:To investigate the inhibitory effect of combined strategy of poly adenosine diphosphate ribose polymerase (PARP) inhibitor and interleukin-1β (IL-1β) inhibitor on homologous recombination deficiency (HRD)-proficient ovarian cancer cells.Methods:(1) HRD-proficient ovarian cancer cell lines OVCAR3 and CAOV3 were treated with PARP inhibitor olaparib. Screening by RNA sequencing analysis, the expression level of IL-1β was validated by enzyme-linked immunosorbent assay (ELISA) and western blot. (2) The dose-response curves of IL-1β inhibitor diacerein were evaluated by cell counting kit-8 (CCK-8) assays in OVCAR3 and CAOV3 cells. CCK-8 assays were further applied to determine the viabilities of OVCAR3 and CAOV3 cells. (3) To evaluate the synergistic effects of olaparib and IL-1β inhibitor in vivo, the transplanted ovarian cancer model was constructed. BALB/c-nude mice ( n=16) were injected intraperitoneally with 1×10 7 OVACR3 cells labelled with luciferase (OVCAR3-Luc). Immunohistochemistry (IHC) assay was performed to determine nuclear antigen associated with cell proliferation (Ki-67) expression. (4) Blood routine tests, kidney and liver function tests were performed to analyze the toxic reaction of different drug treatments. The potential drug-induced injuries of vital organs including heart, liver, spleen, lungs and kidneys of nude mice were determined by hematoxylin-eosin (HE) staining. Results:(1) The RNA sequencing results showed that the mRNA level of IL-1β was the most significantly increased among the 25 differentially expressed genes in OVCAR3 cells treated with olaparib, compared to the negative control group. Olaparib treatment significantly promoted the secretion and expression of IL-1β protein in both OVACR3 and CAOV3 cells by ELISA [(36.2±3.5) and (49.5±3.5) pg/ml, respectively; all P<0.001] and western bolt (2.87±0.37 and 2.05±0.08, respectively; all P<0.01). (2) The half maximal inhibitory concentration (IC 50) value of IL-1β inhibitor was determined as follows: 75 μmol/L for OVACR3 cells and 100 μmol/L for CAOV3 cells. The treatments were divided into four groups including control group, olaparib monotherapy group, IL-1β inhibitor monotherapy group and the combination therapy group. The cell viabilities of each group in OVCAR3 and CAOV3 were determined by CCK-8 assay. The data in each group were showed as follows for OVCAR3 and CAOV3 cells: (100.0±0.4)% and (100.0±3.5)% in control group; (63.1±6.2)% and (63.3±3.8)% in olaparib monotherapy group; (61.6±4.7)% and (63.8±3.5)% in IL-1β inhibitor monotherapy group; and (32.9±5.2)% and (30.0±1.3)% in the combination therapy group. The viability assay showed that the combined strategy exhibited a significant inhibition effect on OVACR3 and CAOV3 cells, compared to the monotherapy group and the control group (all P<0.01). (3) All mice with transplanted tumors of HRD-proficient ovarian cancer cells were randomly divided into four groups, and treated with four different treatments as mentioned above, respectively. After 4 weeks (on day 29), the vivo fluorescence imaging were determined. The results showed that the amount of fluorescence of transplanted tumors was mostly decreased in the combination therapy group [(0.5±0.4)×10 10 p/s], compared to the control group [(4.2±1.0)×10 10 p/s] or the groups treated with any single drug [(3.1±0.9)×10 10, (2.2±0.9)×10 10 p/s; all P<0.05]. Mice were then sacrificed under anesthesia, and all transplanted tumors detached and weighed for further investigation. The weight of transplanted tumors was significantly decreased in the combination therapy group [(0.09±0.03) g], compared to that in control group [(0.25±0.05) g] or groups treated with any single drug [(0.17±0.03), (0.19±0.04) g; all P<0.05]. The measurement of the expression of Ki-67 showed that it was significantly decreased in the combination therapy group (0.33±0.10), compared to that in the control group (1.00±0.20) or monotherapy groups (0.76±0.07, 0.77±0.12; all P<0.05). (4) There were no significant differences of body weights, blood routine test, renal and liver function tests among mice with different treatments (all P>0.05). Moreover, no significant injuries were observed in the vital organs among the four groups. Conclusions:The combination of olaparib and IL-1β inhibitor synergistically exhibits significant cytotoxicity in HRD-proficient ovarian cancer cells. Moreover, the blood routine and blood biochemistry results confirmed the biosafety of the combination of olaparib and IL-1β inhibitor.

Factors affecting long-term survival of advanced high-grade serous ovarian cancer / 中华妇产科杂志

Objective:To identify the factors associated with long-term survival and guide the decision for primary surgery in patients with advanced high-grade serous ovarian cancer(HGSOC).Methods:In this case-control study, clinical parameters, including surgical and non-surgical associated factors, were collected and compared between the patients with short-term (<2 years) and long-term (>5 years) survival who all underwent primary debulking surgery (PDS) followed by carboplatin and paclitaxel chemotherapy from January 2004 to December 2016. Univariate analysis was examined by chi-square test and multivariate analysis was performed by logistic regression analysis.Results:There were 95 cases long-term survival (LTS group) and 77 cases short-term survival (STS group) in 698 newly diagnosed HGSOC patients with International Federation of Gynecology and Obstetrics (FIGO) stage Ⅲc and Ⅳ who met include and exclude criteria. (1) Univariate analysis showed that the proportion of complete cytoreduction with no visible residual disease (R0) at PDS and platinum sensitivity in LTS group were significantly higher than those in STS group ( P<0.01). The surgical complexity score (SCS), the preoperative serum CA 125 level and the ascites volume in the LTS group were significantly lower than those of the STS group (all P<0.05). In the LTS group, the preoperative incidence of lesions in retrograde peritoneum of the bladder, serosal and mesangial membrane of the small intestine, upper abdominal peritoneum and liver parenchyma were significantly lower than those in the STS group (all P<0.05). Multivariate logistic regression analysis showed that platinum sensitivity ( OR=0.016, 95% CI: 0.004-0.063, P<0.01), ascites volume >500 ml ( OR=3.193, 95% CI: 1.285-7.930, P=0.012), and SCS ≥8 ( OR=17.433, 95% CI: 2.281-133.25, P=0.003) were independent factors affecting long-term survival ( P>0.05). (2) Totally 37 of 95 in long-term survival and 16 of 77 in short-term survival achieved R0 cytoreduction at PDS. Univariate analysis showed that preoperative serum CA 125 level, preoperative lesion score, preoperative lesion (DS) score, ascites volume, platinum sensitivity,and SCS were significantly correlated with the R0 PDS (all P<0.05). Multivariate analysis showed that ascites volume >500 ml ( OR=5.199, 95% CI: 2.015-13.409, P=0.001), DS >2 ( OR=15.264, 95% CI: 5.843-39.874, P<0.01) and SCS ≥4 ( OR=4.176, 95% CI: 1.618-10.777, P=0.003) were independent factors associated with R0 cytoreduction. In patients with DS ≤2 or SCS <4, but not those with DS >2 or SCS ≥4, R0 cytoreduction was significantly associated with long-term survival. Conclusion:The intrinsic biology of tumor is the factor influencing long-term survival of advanced HGSOC patients, and those who present with wide intraperitoneal metastases and need to remove multiple organs may not benefit from R0 cytoreduction.

Research progress of Bub1 in cancers / 国际肿瘤学杂志

Bub1 is essential for assembling of the functional spindle assembly checkpoint (SAC) to guarantee the correct separation of sister chromatids. Abnormal expression of Bub1 can elevate defects in SAC function, chromosome instability, and the incidence of aneuploidy. Several recent studies indicate that aberrantly expressed Bub1 may promote the tumorigenesis via regulating cell proliferation, invasion, migration and cancer stem cell activation. Further understanding of the mechanism of Bub1 in malignancy may provide new targeted therapy strategies.

Research advances on the role of exosomes in chemotherapy resistance of ovarian cancer / 浙江大学学报·医学版

Chemotherapy resistance is one of the biggest challenges in treatment of ovarian cancer. Mounting evidence shows that the exosomes shedding from tumor cells are considered to be involved in chemotherapy resistance of ovarian cancer by enhanced exosomal export of drugs, transferring RNAs or proteins and interfering with the bioactivity of therapeutic anti-tumor antibodies. In this review, we display the correlation between exosomes and chemotherapy resistance of ovarian cancer, the mechanism of exosomes involved in chemotherapy resistance of ovarian cancer, and discuss the potential clinical values of exosomes in chemotherapy resistance of ovarian cancer.

Key anatomies of DeLancey's three levels of vaginal support theory: an observation in laparoscopic surgery / 浙江大学学报·医学版

OBJECTIVE@#To observe and verify the key anatomies of DeLancey's three levels of vaginal support theory through laparoscopic surgery by space dissection technique.@*METHODS@#The features and stress performance of related anatomies were observed and analyzed in laparoscopic type C hysterectomy and pelvic lymphadenectomy for cervical cancer by natural space exposures.@*RESULTS@#The main ligament-like structure at level Ⅰ was the uterosacral ligament, which acted as the main apical fixation in the sacral direction, while the cardinal ligament was mainly composed of vascular system, lymph-vessels and loose connective tissue around them, lacking the tough connective tissue structures, which was connected to the internal iliac vascular system. There were no strong ligaments connected to the tendinous arch of pelvic fascia (ATFP) at the lateral side of vaginal wall at level Ⅱ. ATFP was the edge of the superior fascia of pelvic diaphragm, which was bounded by the fascia of the obturator. Its surface was smooth and close to the levator ani muscle and fuses with the vaginal fascia in about one thirds of middle lower segments of the vagina. When the ureter tunnel is separated, dense connective structures can be found in both anterior and posterior walls near the intersection of the ureter across uterine artery, fixing the bilateral angle of the bladder triangle, starting from the cervix and vagina and ending in the tunica muscularis vesicae urinariae.@*CONCLUSIONS@#Based on the laparoscopic anatomy, the pelvic floor fascia ligament support above the levator ani muscle can be considered mainly around the vagina, and fascial ligament above the levator ani muscle can be simply considered as two parallel planes forming a "double hammock" structure, which may provide more anatomic data for pelvic floor reconstruction.

Establishment of a prognostic model for preterm delivery in women after cervical conization / 浙江大学学报·医学版

OBJECTIVE@#To establish a prognostic model for preterm birth in women after cervical conization, and to evaluate its effectiveness.@*METHODS@#Seventy three women after cervical conization in Women's Hospital of Zhejiang University were included for this retrospective study. The influencing factors of preterm delivery were analyzed by Logistic regression analysis and a prognostic model was created. Receiver operating characteristic (ROC) curve was used for evaluation of the predictive ability of the model. Forty five women who underwent cervical conization were included for testing the validity of the model.@*RESULTS@#For women after cervical conization, mother's age (=1.20, 95%:1.01-1.43, <0.05) and cervical length during middle pregnancy (=0.06, 95%:0.01-0.21,<0.01) were independent predictors for preterm birth. The regression model was Logit ()=1.408-2.903×cervical length+0.186×age. The areas under the ROC curve (AUC) of the training dataset was 0.93 (95%:0.87-0.99). The sensitivity, specificity, Youden index, positive predictive value (PPV), negative predictive value (NPV) and accuracy with the cutoff value of -1.512 were 91.7%, 81.5%, 0.732, 68.8%, 95.7% and 84.5% respectively. The AUC of the testing dataset was 0.94 (95%:0.86-1.00). The sensitivity, specificity, Youden index, PPV, NPV and accuracy with the cutoff value of -0.099 were 92.9%, 90.3%, 0.832, 81.3%, 96.5% and 91.1%, respectively.@*CONCLUSIONS@#The model based on the age and cervical length during middle pregnancy can effectively predict preterm delivery in pregnant women after cervical conization.

EMA/CO regimen for chemotherapy 24 patients with ultra high-risk gestational trophoblastic neoplasia / 中华妇产科杂志

Objective To evaluate etoposide,methotrexate and dactinomycin (EMA)/ cyclophosphamide and vincristine (CO) regimen for treatment of ultra high-risk gestational trophoblastic neoplasia (GTN).Methods A total of twenty-four ultra high-risk patients who had International Federation of Gynecology and Obstetrics (FIGO) prognostic scores greater or equal to 12 with liver,brain,or extensive metastases did poorly when treated with primary chemotherapy admitted in Women's Hospital,School of Medicine,Zhejiang University from January 2001 to December 2015.All of the patients were treated by EMA/CO regimen and followed up to death or December 2017.The clinical data of patients were analyzed retrospectively and the efficacy and toxicity of EMA/CO were evaluated.Results All of the cases with ultra high-risk GTN had FIGO prognostic scores ≥12 (ranged 12-18,median 13.0).Twenty patients (83％,20/24) received EMA/CO regimen as primary treatment and 4 patients (17％,4/24) had a history of failed chemotherapy.Seven patients (29％,7/24) had metastasis of liver or brain and 17 patients (71％,20/24)had no metastasis of liver and brain.Twenty-four patients received totally 167 courses of EMA/CO regimen (average 7.0 courses).Sixteen patients achieved complete remission and 8 patients showed drug-resistant.The complete remission rate was 67％ (16/24) and the resistance rate was 33％ (8/24).Of the 16 patients who got complete remission,6 cases were treated with EMA/CO regimen alone,and 10 cases were treated by chemotherapy combined with surgery.For the 8 patients who showed drug-resistant to EMA/CO,5 cases of them received EMA/etoposide and cisplatin (EP) regimen and 3 cases got remission,1 case received methotrexate,dactinomycin and cyclophosphamide (MAC) regimen and got remission,2 cases gave up treatment because of economic factors.The side effects of EMA/CO mainly included Ⅲ-Ⅳ degree neutropenia,anemia and alopecia.The incidence of Ⅲ-Ⅳ degree neutropenia during the treatment of EMA/CO was 21.6％ (36/167),the incidence of anemia was 96.4％ (161/167),and the incidence of alopecia was 60.5％ (101/167).In these 24 ultra high-risk GTN patients,4 patients died during follow-up.In the 20 patients who got complete remission,no recurrence or secondary tumor by chemotherapy were occurred.Conclusion EMA/CO is an effective regimen with manageable toxicity for patients with ultra high-risk GTN.

Magnetic resonance imaging for pelvic tumors during pregnancy: manifestations and clinical application / 中华围产医学杂志

Objective To investigate the manifestations and clinical application of magnetic resonance imaging (MRI) in the diagnosis of pelvic tumors during pregnancy.Methods Clinical data and MRI manifestations of 161 patients attending Women's Hospital,School of Medicine,Zhejiang University with pelvic tumors during pregnancy between March 2013 and July 2016 were retrospectively analyzed.MRI manifestations and clinical treatments of different tumors in pregnancy were summarized.All data were analyzed by descriptive statistics.Results (1) The gestational ages at initial diagnosis of the 161 patients ranged between 8-39 weeks and the average gestational age was (29±-4) weeks.Of all patients,147 (91.3％) had benign tumor,while the other 14(8.7％) were diagnosed as malignanices.The sensitivity,specificity and accuracy of MRI in the diagnosis of malignancy during pregnancy were 92.8％ (13/14),97.3％ (143/147) and 96.7％ (156/161).Uterine neoplasm,ovarian tumor,intestinal tumor and pelvic peripheral nerve schwannoma respectively accounted for 67.1％ (108/161),29.8％ (48/161),1.9％ (3/161) and 1.2％ (2/161) of all cases.(2) Among the 108 gravidas with uterine neoplasm,104 cases of leiomyoma continued to term pregnancy,among which 71 underwent myomectomy after vaginal delivery and 33 had myomectomy during cesarean section;the other four cases of cervical cancer terminated their pregnancy.(3) Among the 48 gravidas with ovarian tumor,38 were diagnosed as having benign tumor by MRI and continued to term pregnancy;five were diagnosed with tumor pedicle torsion and treated with emergency operation because of acute abdominal disease;two patients who were both in their second trimester (16 and 20 gestational weeks) were diagnosed with ovarian cancer complicated with peritoneal metastasis by MRI and decided to terminate pregnancy;another two patients with uncertain preoperative diagnosis of ovarian cancer were followed up closely to term and the tumor was removed during cesarean section;one patient with ovarian malignant tumor,which was diagnosed by MRI at the 32nd gestational week,received cesarean section and cytoreductive surgery.(4) In the three gravidas with intestinal tumor,one was complicated with sigmoid colon cancer and underwent cesarean section,radical resection of sigmoid colon tumor,colostomy and enterodialysis at 28+3 weeks of gestation;one with appendiceal mucinous adenocarcinoma was monitored closely until full-term pregnancy and received right hemicolectomy during cesarean section;the third case with malignant stromal tumor of the small intestine underwent cesarean section at 30+5 weeks of gestation as well as the completer resection of tumor.(5) One gravida with pelvic peripheral nerve schwannomas was diagnosed with pelvic schwannoma and underwent cesarean section and tumor extirpation at term.The other one underwent tumor extirpation prior to delivery due to the larger size of tumor and had a vaginal delivery at term.Conclusions MRI can provide more precise information for prenatal diagnosis of pelvic tumors during pregnancy.

Expression of miR-let-7e-3p in cervical intraepithelial neoplasm and cervix carcinoma and its clinical significance / 浙江大学学报·医学版

To investigate the expression of microRNA (miRNA, miR) let-7e-3p in different cervical lesions and its clinical significance.The expression of miR-let-7e-3p in the tissues of normal cervix (=26), high-grade squamous intraepithelial lesion (HSIL) (=37), and cervix carcinoma (=101) were detected by reverse transcription and quantitative polymerase chain reaction (RT-qPCR). The correlation of miR-let-7e-3p expression with the clinicopathological parameters of patients with cervical cancer was analyzed. miR-let-7e-3p mimic was transfected into cervical carcinoma Siha cells. The cell cycle and apoptosis were determined by flow cytometry; cell proliferation was determined by CCK-8 kit; and the migration and invasion of cells were determined by Transwell assay.The relative expression levels of miR-let-7e-3p in normal cervix, HSIL, and cervical carcinoma were 1.45±0.24, 0.79±0.05 and 0.46±0.04, respectively (all<0.05). After transfection with miR-let-7e-3p mimic, the S-phase fraction and apoptosis rate of Siha cells were increased significantly compared with control group[(29.76±6.6)% vs (13.38±1.3)%,<0.05; (5.98±1.38)% vs (3.53±0.79)%,<0.05, respectively]. OD of transfected Siha cells at 48, 72 and 96 h were 0.57±0.11,0.65±0.04 and 0.84±0.14, which were significantly lower than those of untransfected Siha cells (0.74±0.05, 0.93±0.10 and 1.47±0.14, all<0.05). The migration and invasion abilities of transfected Siha cells were not significantly changed (all>0.05).The expression of miR-let-7e-3p is down-regulated in cervical neoplasms, which is associated with cell cycle arrest and proliferation inhibition of cervical cancer cells.

Clinical significance of HPV L1 capsid protein detection in cervical exfoliated cells in high-risk HPV positive women / 中华妇产科杂志

Objective To explore the clinical significance of human papillomavirus L1 capsid protein detection in cervical exfoliated cells in high-risk HPV positive women. Methods From November 2012 to June 2013,386 high-risk HPV positive (detected by hybrid capture Ⅱ) cases were enrolled as eligible women from Huzhou Maternity&Child Care Hospital and Women′s Hospital,School of Medicine, Zhejiang University. All eligible women underwent liquid-based cytology (ThinPrep) followed by colposcopy. Biopsies were taken if indicated. Cervical exfoliated cells were collected for HPV L1 capsid protein detection by immunocytochemistry. Expression of HPV L1 capsid protein in groups with different histological diagnosis were compared, and the role of HPV L1 capsid protein detection in cervical exfoliated cells in cervical lesions screening was accessed. Results Total 386 enrolled eligible women were finally diagnosed histologically as follwed:162 normal cervix, 94 low-grade squamous intraepithelial lesion (LSIL), 128 high-grade squamous intraepithelial lesion (HSIL) and 2 squamous cervical cancer (SCC). The positive expression rate of HPV L1 in HSIL+(HSIL or worse) group was significantly lower than that in LSIL-(LSIL or better) group (19.2% vs 66.4%,P=0.000). While identifying HSIL+ in HPV positive cases and compared with cytology, HPV L1 detection resulted in significant higher sensitivity (80.77%vs 50.77%,P=0.000) and negative predictive value (NPV;87.18% vs 76.47%,P=0.004), significant lower specificity (66.41% vs 81.25%,P=0.000),and comparable positive predictive value (PPV;54.97% vs 57.89%, P=0.619). To identify HSIL+in HPV-positive/cytology-negative women, the sensitivity, specificity, PPV, and NPV of HPV L1 detection were 87.50%, 61.54%, 41.18%, and 94.12%respectively, while 80.00%, 86.36%, 80.00%and 86.36%respectively in HPV-positive/atypical squamous cell of undetermined significance(ASCUS)women. Conclusions HPV L1 capsid detection in cervical exfoliated cells have a role in cervical lesions screening in high-risk HPV positive women, and may be a promising triage for high-risk HPV-positive/cytology-negative or ASCUS women.