Development and Validation of a Novel Gene Signature for Predicting the Prognosis by Identifying m5C Modification Subtypes of Cervical Cancer.

Background: 5-Methylcytidine (m5C) is the most common RNA modification and plays an important role in multiple tumors including cervical cancer (CC). We aimed to develop a novel gene signature by identifying m5C modification subtypes of CC to better predict the prognosis of patients. Methods: We obtained the expression of 13 m5C regulatory factors from The Cancer Genome Atlas (TCGA all set, 257 patients) to determine m5C modification subtypes by the "nonnegative matrix factorization" (NMF). Then the "limma" package was used to identify differentially expressed genes (DEGs) between different subtypes. According to these DEGs, we performed Cox regression and Kaplan-Meier (KM) survival analysis to establish a novel gene signature in TCGA training set (128 patients). We also verified the risk prediction effect of gene signature in TCGA test set (129 patients), TCGA all set (257 patients) and GSE44001 (300 patients). Furthermore, a nomogram including this gene signature and clinicopathological parameters was established to predict the individual survival rate. Finally, the expression and function of these signature genes were explored by qRT-PCR, immunohistochemistry (IHC) and proliferation, colony formation, migration and invasion assays. Results: Based on consistent clustering of 13 m5C-modified genes, CC was divided into two subtypes (C1 and C2) and the C1 subtype had a worse prognosis. The 4-gene signature comprising FNDC3A, VEGFA, OPN3 and CPE was constructed. In TCGA training set and three validation sets, we found the prognosis of patients in the low-risk group was much better than that in the high-risk group. A nomogram incorporating the gene signature and T stage was constructed, and the calibration plot suggested that it could accurately predict the survival rate. The expression levels of FNDC3A, VEGFA, OPN3 and CPE were all high in cervical cancer tissues. Downregulation of FNDC3A, VEGFA or CPE expression suppressed the proliferation, migration and invasion of SiHa cells. Conclusions: Two m5C modification subtypes of CC were identified and then a 4-gene signature was established, which provide new feasible methods for clinical risk assessment and targeted therapies for CC.

[Prognostic analysis of radical radiotherapy in stage Ib and IIa cervical carcinoma].

OBJECTIVE: To investigate the efficacy and prognostic factors in patients with stage Ib and IIa cervical carcinoma by radical radiotherapy. METHODS: Between January 1999 and January 2012, 108 patients with stage Ib and IIa cervical carcinoma received radical radiotherapy in Cancer Hospital, Chinese Academy of Medical Sciences were included and analyzed retrospectively. Patients of stage Ib1, Ib2, IIa1 and IIa2 were 18 (16.7%, 18/108), 38 (35.2%, 38/108), 33 (30.6%, 33/108) and 19 (17.6%, 19/108), respectively. RESULTS: The 5-year overall survival rate was 76.2% and the 5-year disease free survival rate was 75.6%. Totally 25 (23.1%, 25/108) patients developed recurrent disease, 16 of them (64%, 16/25) had local recurrences, 6 (24%, 6/25) had distant metastases and 3 cases had both local recurrence and distant metastases. Among patients with recurrent disease, 23 died and 2 survive with tumor. Totally 24 patients died, 23 of them died due to tumor recurrence and the other one died of other reason. The univariate analysis showed that, lymph node metastasis, squamous cell carcinoma antigen (SCC) levels before treatment, SCC levels after treatment 1 month had relation with overall survival time in patients with stage Ib and IIa cervical carcinoma (all P < 0.05). The multivariate analysis showed that, lymph node metastasis and SCC levels after treatment 1 month were the independent prognostic factors for overall survival time for the cervical squamous cell carcinoma (OR = 2.5, 4.4; all P < 0.05). CONCLUSIONS: By means of radical radiotherapy, stageIb and IIa cervical carcinoma patients with lymph node metastasis and SCC levels ≥ 1.5 mg/L after treatment one month have poor prognosis.While, stageIb and IIa patients with concurrent chemoradiotherapy after neoadjuvant chemotherapy did not affect the prognosis. The 5-year survival rate with concurrent chemoradiotherapy was higher than that of radiotherapy.

[Study of the radiotherapy modality for patients with stage IIb-IIIb cervical stump cancer].

OBJECTIVE: To investigate the radiotherapy modality progress of stageIIb-IIIb cervical stump cancer. METHODS: The clinical data of 13 patients with stageIIb-IIIb cervical stump cancer undergoing radiotherapy from January 2000 to April 2012 was reviewed. Before 2006, 8 patients received conventional external beam radiotherapy and brachytherapy.Since 2006, 5 patients received intensity-modulated radiotherapy (IMRT) and brachytherapy. RESULTS: The median survival was 12-139 months. The median overall survivals and disease free survivals in the conventional radiotherapy (CRT) group were 57 months and 50 months, 3 cases of them recurred during 8-19 months and died of tumor progression.While, the median overall survivals and disease free survival in the IMRT group both were 21 months and nobody recurred. In the CRT group, 7 patients suffered toxicities, including 5 patients grade I-II acute rectum reaction, 2 patients grade I bladder reaction; and 3 had grade I-III, late rectum reaction, 2 patients for grade II bladder late reaction.In the IMRT group, toxicities including 1 case grade I acute or late rectum reaction, and no bladder reaction. CONCLUSION: In our experience, the recommended IMRT and interstitial brachytherapy for the selected patients with advanced cervical stump carcinoma may be obtain better tumor dose distribution and more sparing of the organ at risk.

A preliminary study of genes related to concomitant chemoradiotherapy resistance in advanced uterine cervical squamous cell carcinoma.

BACKGROUND: Tumor intrinsic chemoradiotherapy resistance is the primary factor in concomitant chemoradiotherapy failure in advanced uterine cervical squamous cell carcinoma. This study aims to identify a set of genes and molecular pathways related to this condition. METHODS: Forty patients with uterine cervical squamous cell carcinoma in International Federation of Gynecology and Obstetrics stage IIb or IIIb, treated with platinum-based concomitant chemoradiotherapy between May 2007 and December 2012, were enrolled in this trial. Patients included chemoradiotherapy resistant (n = 20) and sensitive (n = 20) groups. Total RNA was extracted from fresh tumor tissues obtained by biopsy before treatment and microarray analysis was performed to identify genes differentially expressed between the two groups. RESULTS: Microarray analysis identified 108 genes differentially expressed between concomitant chemoradiotherapy resistant and sensitive patients. Functional pathway cluster analysis of these genes revealed that DNA damage repair, apoptosis, cell cycle, Map kinase signal transduction, anaerobic glycolysis and glutathione metabolism were the most relevant pathways. Platelet-derived growth factor receptor alpha (PDGFRA) and protein kinase A type 1A (PRKAR1A) were significantly upregulated in the chemoradiosensitive group, while lactate dehydrogenase A (LDHA), bcl2 antagonist/killer 1 (BAK1), bcl2/adenovirus E1B 19 kDa interacting protein 3 (BNIP3), single-strand-selective monofunctional uracil-DNA glycosylase 1 (SMUG1), and cyclin-dependent kinase 7 (CDK7) were upregulated in the chemoradiotherapy resistant group. CONCLUSION: We have identified seven genes that are differentially expressed in concomitant chemoradiotherapy resistant and sensitive uterine cervical squamous cell carcinomas, which may represent primary predictors for this condition.

[Clinical analysis of 44 cases with malignant transformation of ovarian mature cystic teratoma].

OBJECTIVE: To analyze the clinicopathologic characteristics, treatment and prognostic factors in malignant transformation of mature cystic teratoma (MCT) of ovary. METHODS: The clinical data of 44 patients with MCT from January 1961 to June 2009 were reviewed. RESULTS: The median age of the 44 patients was 48 years (range, 16 - 84 years). Mean tumor size was (16 ± 6) cm. Thirty-two cases were diagnosed squamous cell carcinoma (73%, 32/44), and 5 of them with the elevated level of serumal squamous cell antigen (SCC-Ag). Three of 37 cases (8%, 3/37) were identified with malignant transformation in image examinations. Rapid frozen section examination and multiple-location biopsy were performed in 8 cases, and 5 of them were detected with malignant diseases. Twenty-two patients with disease confined within the unilateral ovary (10 with intact capsule, and 12 with ruptured capsule). Diseases extended extra ovaries in the others 22 patients. The median cumulative overall survivals were 126 and 10 months, respectively. The difference between the two groups was significant (P < 0.01). Twenty-seven patients had no residual tumor after primary surgery. The median cumulative overall survivals between the patients with and without residual tumor were 10 and 84 months respectively, and there were significant difference between two groups (P < 0.01). Seven selected patients with malignant disease confined within unilateral ovary underwent fertility-sparing surgery, and 2 cases of them had successful pregnancies and delivery, while other 4 cases with ruptured capsule recurred. CONCLUSIONS: The most common pathology type of malignant transformation in mature cystic teratoma of the ovary is squamous cell carcinoma. Comprehensive pre-operation image examination and tumor marker level detection might be of great help in diagnosis. Tumor extension extraovary and residual tumor after surgery are the most significant poor prognostic factors. Early stage patient with ruptured capsule should be very discreet to choose fertility-sparing surgery.

[Clinical review of 97 patients with endometrial stromal sarcoma].

OBJECTIVE: To review the survival outcomes in patients with endometrial stromal sarcoma (ESS) in Cancer Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, and to discuss prognostic factors and the role of post-operative adjuvant radiotherapy and chemotherapy. METHODS: Hospital records and pathology reports for 97 patients with ESS were reviewed. Among 97 patients, 69 had low-grade ESS (LGESS), 16 had high-grade ESS (HGESS) and 12 had unclear grade. The median age at diagnosis was 44.0 years. The median follow-up time was 62 months (5 - 277 months). Atypical vaginal bleeding (43%) and prolonged and increased menses (36%) were the main symptoms. RESULTS: Totally 2-year and 5-year cumulative survival rates were 93% and 84%, respectively. Cumulative survival curves were significantly different between LGESS and HGESS, and so did cumulative survival curves between stage I - II and stage III - IV (P < 0.05). Totally, 34 patients (37%) had local or distant recurrence. The median time-to-recurrence (TTR) was 27 months. The recurrence rates of the patients with or without preserve of ovary were 89% and 24%, respectively (P = 0.000). The local-control-rates of the patients who received or did not receive post-operative radiotherapy were 81% and 43%, respectively (P = 0.011). CONCLUSIONS: The prognosis of HGESS is obviously worse than that of LGESS. The risk of recurrence of patients with preserve of ovary was remarkably higher than that of patients without preserve of ovary. Postoperative radiotherapy could increase the local-control-rates.

Treatment options in stage I endometrial stromal sarcoma: a retrospective analysis of 53 cases.

OBJECTIVE: To discuss the optimal treatment options for stage I patients with endometrial stromal sarcoma (ESS). METHODS: We reviewed hospital records and pathology of 53 patients with ESS at stage I. Statistical analysis was performed using SPSS 12.0 software, and Chi-square test, t-test and log rank test were adopted. RESULTS: Among 53 patients, 37 had low-grade tumors, 11 had undifferentiated endometrial sarcoma (UES) and 5 had unclassified ESS. The median follow-up time was 66 months, and 48 cases were still alive. The overall 2-year and 5-year survival rates were 91.5% and 85.9%, respectively. The recurrence rate of the patients with preserved ovarian function was remarkably higher than that of patients without (100% vs. 22.7%, P<0.001). The patients who received adjuvant whole pelvic radiation (Dt 40 approximately 45 Gy) had obviously higher local control rate than the patients who did not (93.8% vs. 57.1%, P=0.007), but they had similar survival (P=0.963). Among 7 of the 11 UES patients without distant recurrence, 5 received the adjuvant chemotherapy with IAP (ifosfamide 1.0 g, d1-4; epirubicin 25 mg/m2, d1-2; cisplatin 20 mg, d1-5; mensa 0.2 g, 0, 4, 8 h from the application of ifosfamide, d1-4, q 28 days) or VAD (vincristine 1.2 mg/m2, d1; adriamycin 20 mg/m2, d1-3; dacarbazine 250 mg/m2, d1-5, q 28 days), and none of the other 4 cases recurring distantly received the chemotherapy with IAP or VAD. CONCLUSIONS: Surgeries not preserving ovarian function were helpful to decrease the risk of recurrence compared with those surgeries sparing ovarian function. Adjuvant radiotherapy could improve local control but not survival. Adjuvant chemotherapy with IAP or VAD seemed to be beneficial to UES patients.

[Clinical analysis of 42 cases of primary malignant melanoma in female genital tract].

OBJECTIVE: To analyze the clinical characteristics, diagnosis, treatment and prognosis of primary malignant melanoma in female genital tract. METHODS: The clinical data of 42 patients of primary malignant melanoma in female genital tract were reviewed. RESULTS: The tumors were originated from vulva, vagina and cervix in 14 (33%), 23 (55%) and 5 (12%) cases, respectively. Thirty-eight cases had biopsies. Among them, 6 cases were misdiagnosed. Eighteen surgical specimens were examined by immunohistochemistry assays. S-100 protein was positive in all cases, and monoclonal antibody to melanoma of human (HMB-45) was positive in 14 cases. The 2-year and 5-year cumulative recurrence-free survival rates were 35% and 23% respectively, while the 2-year and 5-year cumulative overall survival rates were 53% and 27% respectively. The 2-year cumulative overall survival rates for the patients of early stage [International Federation of Gynecology and Obstetrics (FIGO) stage I and II] and that of advanced stage (stage III and IV) were 77% and 34% respectively (P < 0.05). The 2-year cumulative overall survival rates for the patients of stage I and stage II were 78% and 74% respectively (P = 0.303). In the 40 patients who received surgery, univariate analysis showed that the adjuvant chemotherapy improved the recurrence-free survival and the overall survival significantly (P < 0.05), and the other factors including radical surgery, regional lymphadenectomy, biotherapy and radiotherapy did not affect prognosis (P > 0.05). Compared with chemotherapy, biochemotherapy did not improve prognosis significantly (P > 0.05). CONCLUSIONS: Biopsy for the malignant melanoma in female genital tract has high misdiagnosis rate. Immunohistochemistry assay could improve diagnosis markedly. The FIGO staging system fails to predict the prognosis accurately. Surgery plays an important role in treatment, while the adjuvant chemotherapy could improve survival effectively.