Case report: Squamous cell carcinoma and spindle cell sarcoma (SCS) arising in a mature cystic teratoma of the ovary.

Objective: Malignant transformation of mature ovarian teratoma is a rare phenomenon, mainly occurring in postmenopausal period. Squamous cell carcinoma accounts for 80% of all malignant transformations. Sarcoma transformation is much less common and tends to imply a poorer prognosis and aggressiveness. Case report: We report a case of undifferentiated sarcoma with squamous cell carcinoma in a mature cystic teratoma of the ovary in a 36-year-old woman. The tumor shows epithelial and stromal components. This is a unique report of a benign teratoma of the ovary with malignant transformation, showing epithelial and sarcomatous components. This young woman presented with abdominal distension and a rapidly enlarging ovario-derived pelvic mass with a slightly elevated CA199 tumor marker of 115.9 U/ml. The woman underwent transabdominal excision of the left ovarian cyst on October 20, 2020. During the operation, rapid freezing pathological examination did not indicate malignancy. The postoperative paraffin pathology revealed undifferentiated sarcoma with squamous cell carcinoma (from mature cystic teratoma malignancy), and she finally received comprehensive staging surgery. Postoperative paraffin pathology showed no residual cancer in uterus and other tissues, and all lymph nodes were negative. The patient was finally diagnosed with ovarian malignant tumor IC1 stage (high-grade spindle cell sarcoma complicated with squamous cell carcinoma). Chemotherapy was completed three times after surgery, and no signs of recurrence were found after follow-up. Conclusion: The preoperative diagnosis and intraoperative rapid freezing examination of malignant transformation of mature teratoma of ovary are challenging.

[Study on high risk factors associated with positive margin of cervix conization in patient with cervical intraepithelial neoplasia].

OBJECTIVE: To assess the high risk factors associated with the positive margin of conization in patients with cervical intraepithelial neoplasia (CIN). METHODS: From January 2000 to February 2008, 1699 consecutive patients with CIN undergoing conization was reviewed retrospectively in order to analyze the relationship between the positive margin of conization with clinical prognostic factors, such as patients age, disease grade, size of lesion, the procedure of excision and menopause. chi2 tests was used to compare the different frequencies of factors in groups of positive and negative margin conization, then seven factors with positive margin were processed into unconditional logistic regression analysis. RESULTS: The rate of the positive margin in 1699 patients was 14.01% (238/1699). The mean age of patients with positive margins was (39+/-9) years old, while patients with negative margins was (39+/-8) years old, which didn't reach statistical difference (P>0.05). The rate of the positive margin was 8.63% in cold knife cone (CKC) and 18.66% in loop electrosurgical excision procedure (LEEP), which showed significant difference (P<0.01). Among 1699 patients, 90 patients were with CINI, 339 patients were with CIN II, 1113 patients were with CIN III [including 972 with severe dysplasia and 141 with cancer in situ (CIS)], 87 patients were with cervical cancer stage Ia1, 70 patients were with stage Ia2 or advanced stages. The rate of positive margin was 1.11% (1/90), 3.83% (13/339), 10.70% (104/972), 26.24% (37/141), 35.63% (31/87) and 74.29% (52/70), respectively. There was statistic difference among them, except CINI and CINII. When combined CIN I with CIN II, then compared with CIN III, cervical cancer with Ia1 and Ia2, it also showed statistical difference (P<0.05). The rate of positive margin in postmenopausal women was 21.54% (28/130), which was significantly higher than 13.38% (210/1569) in premenopausal women (P=0.010). The logistic regression analysis showed that the procedure of excision, grade of disease, size of lesion, surface of cervix, and menopause were high risk factors associated with the positive margin, the risk ratio were 5.147, 3.048, 1.271, 1.905 and 1.860, respectively. CONCLUSIONS: High grade, the extent of CIN disease, LEEP and postmenopausal age are high-risk factors associated with positive margin in patients treated by conization. It should be warranted in those patients when designing conization treatment.

Methotrexate induces apoptosis of human choriocarcinoma cell line JAR via a mitochondrial pathway.

OBJECTIVE: To investigate methotrexate (MTX)-induced apoptosis and the involved pathways in human choriocarcinoma cells. STUDY DESIGN: MTX-induced apoptosis of human choriocarcinoma cell line JAR was examined using a PI/Annexin V stain with flow cytometer (FCM). Mitochondrial apoptosis was detected by fluorescence microscopy, and analyzed by FCM using a MitoCapture mitochondrial apoptosis detection kit. The activities of caspase-8 and caspase-9 were quantified by microtiter plate reader at 405 nm using FLICE/Caspase-8 colorimetric assay kit and Caspase-9/Mch6 colorimetric assay kit. The changes in Bax and Bcl-2 expression were detected during apoptosis using immunocytochemistry and Western blot analysis. RESULTS: JAR cells underwent apoptosis after exposure to 0.1-2.5 microg/ml MTX for 48 h. Decreased mitochondrial membrane potential was observed both by fluorescence microscopy and FCM. The activation of caspase-9 was increased 4.35+/-0.76-fold in MTX-incubated JAR, while there was no obvious change in the activation of caspase-8. When JAR cells underwent apoptosis, the expression of Bcl-2 was decreased and the expression of Bax was increased; both were detected by immunocytochemistry assay. CONCLUSION: Methotrexate in lower concentrations induces apoptosis of human choriocarcinoma cells via mitochondrial-initiated pathways, including reduction of mitochondrial membrane potential, activation of caspase-9, and up-regulation of Bax/Bcl-2 expression.

[Effects of primary chemotherapy with single methotrexate on low-risk gestational trophoblastic neoplasia and influencing factors thereof].

OBJECTIVE: To evaluate the effects of primary chemotherapy with single-agent methotrexate (MTX) on low-risk gestational trophoblastic neoplasia (GTN) and the influencing factors thereof. METHODS: Sixty-one GTN patients with the score of < or = 6 according to the new International Federation of Gynecology and Obstetrics (FIGO) scoring system (2000) were divided into 2 groups: 51 patients were treated with single MTX 0.4 mg/kg daily for 5 days (MTX 5 d group), and 10 patients were treated with MTX on the days 1, 3, 5, and 7, and with folinic acid (FA) 0.1 mg/kg on the days 2, 4, 6, and 8 (MTX + FA group), both group with an interval of treatment course of 2 weeks. The serum level of human chorionic gonadotropin (hCG) was detected every week. If a plateau or increase of serum hCG appeared between 2 examination results, meaning tolerance to MTX, the patients concerned had to undergo different regimens of salvage chemotherapy, all with MTX as one of their components. Univariate and multivariate methods were used to analyze the relationships of different factors to the outcomes of chemotherapy. RESULTS: Thirty-five of the 51 patients of the MTX 5d group (68.6%) achieved complete primary remission, 3 of the 10 patients of the MTX + FA group achieved complete primary remission, and all patients achieved complete remission after salvage chemotherapy. Univariate analysis showed that the mean pretreatment serum level of hCG, duration between antecedent pregnancy and start of treatment, size of tumor, FIGO score, specific regimen of MTX were significantly associated with outcome of chemotherapy (P = 0.004, 0.022, 0.017, 0.005, 0.021 respectively). Logistic regression analysis showed that only three independent factors predictive for the outcome of chemotherapy: MTX regimen (OR = 2.476), FIGO score (OR = 1.431), and pretreatment hCG titer (OR = 1.001). CONCLUSION: Primary chemotherapy with single MTX regimen may still be one of the options for patients with low-risk GTN according to the new FIGO scoring system, though the rate of complete primary remission appears to be lower. All patients with low-risk GTN achieve complete remission after salvage chemotherapy. MTX regimen, FIGO score, and pretreatment hCG are independent risk factors of outcome of single MTX chemotherapy.

[cDNA microarray analysis of gene expression in acquired methotrexate-resistant of human choriocarcinoma].

OBJECTIVE: To identify the molecular components involved in acquired methotrexate-resistance of human choriocarcinoma. METHODS: Methotrexate-resistant cell line (JAR/MTX) was derived from JAR cell line by exposed to intervally and progressively increasing higher concentration of MTX. cDNA microarray analyses of JAR/MTX and its parental cell line JAR were performed. RESULTS: JAR/MTX was established after one year with stable resistance. Its resistant index to MTX was 7.3. Nine genes were differential expressed between JAR/MTX and JAR cells. INSR, SLC1A3, SAT, HBB, and FLJ12443 were underexpressed and HS1, TXNRD1, TAGLN2, and EEF2 were overexpressed in JAR/MTX cells. CONCLUSION: The cDNA microarray system showed that several alterations of gene expression were present in acquired methotrexate-resistance of human choriocarcinoma.

[Establishment of methotrexate-resistant human choriocarcinoma cell line and its biological characteristics].

OBJECTIVE: To establish a methotrexate (MTX)-resistant choriocarcinoma cell line and to determine its biologic characteristics. METHODS: MTX-resistant cell line (JAR/MTX) was derived from human choriocarcinoma cell line JAR by exposed to intermittently and progressively increasing concentration of MTX. Drug sensitivity was detected by MTT; P-gp GST-Pi and PCNA expressions were detected by immunohistochemistry. Cell apoptosis was detected by flow cytometry (FCM) with PI/Annexin V stain. Growth rates and human chorionic gonadotropin (HCG) production were also measured. RESULTS: JAR/MTX cell line was established with stable MTX-resistance (resistance index to MTX was 7.3) and cross-resistant to TAX and VCR. Growthrate of JAR/MTX was lower than that of parent cell line JAR. Expression level of PCNA in JAR/MTX was lower than that in JAR (3.09+/-0.42 compared with 3.72+/-0.35, P<0.05), while GST-pi expression was higher. No statistical difference of P-gp expression existed between two cell lines. JAR/MTX secreted more HCG than JAR every 10(5) cells secreted (95.7+/-5.4 compared with 41.3+/-2.8)mIU after 48 h(P<0.01). The flow cytometry showed that the spontaneous and MTX induced apoptosis in JAR/MTX was significantly lower than that in JAR P<0.05. CONCLUSION: JAR/MTX cell line presented stable resistant to MTX and cross-resistant to TAX and VCR, which might sever as a model in study of drug resistance in choriocarcinoma.

[Efficacy and side effects of methotrexate in the treatment of ectopic pregnancy].

OBJECTIVE: To study the efficacy and side effects of methotrexate with different protocols in the treatment of ectopic pregnancy (EP). METHODS: Total of 648 patients who had EP and were treated only with MTX were analysed. Patients were divided into six groups according to protocol: Group 1, 100 mg intravenously (IV); Group 2, 100 mg, IV, followed by citrovorum factor; Group 3, 20 mg, IV every day for five days; Group 4, 20 mg intramusculary (IM) every day for five days; Group 5, 75 mg, IV; Group 6, 75 mg, IM. RESULTS: The rates of repeated MTX injection in group 1 - 6 because of inadequate decrease of hCG were 23.3%, 25.0%, 21.4%, 20.6%, 24.4% and 22.4% respectively. Success rates were 87.4%, 85.4%, 90.5%, 92.6%, 86.3% and 91.4% respectively. Rates of liver dysfunction were 10.3%, 8.3%, 64.3%, 69.1%, 8.7% and 31.0%. CONCLUSION: Single-dose of 75 mg MTX IV injection may be the best regimen in the treatment of EP because of the same efficacy but the least side effects.