NDR1 increases NOTCH1 signaling activity by impairing Fbw7 mediated NICD degradation to enhance breast cancer stem cell properties.

BACKGROUND: The existence of breast cancer stem cells (BCSCs) causes tumor relapses, metastasis and resistance to conventional therapy in breast cancer. NDR1 kinase, a component of the Hippo pathway, plays important roles in multiple biological processes. However, its role in cancer stem cells has not been explored. The purpose of this study was to investigate the roles of NDR1 in modulating BCSCs. METHODS: The apoptosis was detected by Annexin V/Propidium Iodide staining and analyzed by flow cytometry. BCSCs were detected by CD24/44 or ALDEFLUOR staining and analyzed by flow cytometry. The proliferation ability of BCSCs was evaluated by sphere formation assay. The expression of interested proteins was detected by western blot analysis. The expression of HES-1 and c-MYC was detected by real-time PCR. Notch1 signaling activation was detected by luciferase reporter assay. Protein interaction was evaluated by immunoprecipitation. Protein degradation was evaluated by ubiquitination analysis. The clinical relevance of NDR1 was analyzed by Kaplan-Meier Plotter. RESULTS: NDR1 regulates apoptosis and drug resistance in breast cancer cells. The upregulation of NDR1 increases CD24low/CD44high or ALDEFLUORhigh population and sphere-forming ability in SUM149 and MCF-7 cells, while downregulation of NDR1 induces opposite effects. NDR1 increased the expression of the Notch1 intracellular domain (NICD) and activated the transcription of its downstream target (HES-1 and c-MYC). Critically, both suppression of Notch pathway activation by DAPT treatment or downregulation of Notch1 expression by shRNA reverses NDR1 enhanced BCSC properties. Mechanically, NDR1 interactes with both NICD or Fbw7 in a kinase activity-independent manner. NDR1 reduces the proteolytic turnover of NICD by competing with Fbw7 for NICD binding, thereby leading to Notch pathway activation. Furthermore, NDR1 might function as a hub to modulate IL-6, TNF-α or Wnt3a induced activation of Notch1 signaling pathway and enrichment of breast cancer stem cells. Moreover, we find that the elevation of NDR1 expression predictes poor survival (OS, RFS, DMFS and PPS) in breast cancer. CONCLUSION: Our study revealed a novel function of NDR1 in regulating BCSC properties by activating the Notch pathway. These data might provide a potential strategy for eradicating BCSC to overcome tumor relapses, metastasis and drug resistance.

Notch1 Affects Chemo-resistance Through Regulating Epithelial-Mesenchymal Transition (EMT) in Epithelial Ovarian cancer cells.

Background: Epithelial ovarian cancer (EOC) is the most lethal gynecological malignancy, chemo-resistance is the main cause for treatment failure. Our previous studies have found that SKOV3 could promote immune escape and tumor progression via Notch1 pathway. Therefore, Notch1 is suspected to be involved in chemo-resistance. The current study is to investigate the possible mechanisms of platinum-resistance in epithelial ovarian cancer mediated by Notch1. Methods: The expressions of Notch1, Snail, MMP-2, N-cadherin, Vimentin and E-cadherin were detected by Western-blot. A stable high expression or low expression of Notch1 in ovarian cancer cells was established by using lentiviral gene engineering. The cell migration and invasion ability were observed by scratch test and transwell test. Cell apoptosis rate and cell cycle were analyzed by flow cytometry. Results: The expression levels of Notch1, Snail, MMP-2, N-cadherin and Vimentin in ovarian cancer were high, while the expression levels of E-cadherin were low.Notch1 promoted the expression of Snail, vimentin, N-cadherin and MMP2 protein, but inhibiting the expression of E-cadherin, promoting cell migration and invasion. Notch1 affected apoptosis of cells through Epithelial-Mesenchymal Transition (EMT), increasing the proportion of cells in S phase and G2 phase, thus affecting drug resistance. Conclusion: Notch1 affects EOC cells chemo-resistance by regulating EMT. This may provide a new target for the treatment of ovarian cancer.

Clinical Predictors of Recurrence and Prognostic Value of Lymph Node Involvement in the Serous Borderline Ovarian Tumor.

AIM: This study was aimed to evaluate the risk factors of recurrence and the value of nodal involvement in patients with serous borderline ovarian tumors (SBOT). METHODS: Two hundred twenty-five patients who underwent surgery and were diagnosed with SBOT were retrospectively studied. Univariate and multivariate analyses were used to assess the risk factors for recurrence. Patients' clinical pathologic characteristics were compared between the patients who presented lymph node involvement and those who did not. The significant values of lymph condition influencing 5-year disease-free survival were also evaluated by statistical analysis. RESULTS: Both univariate and multivariate analyses showed that risk factors for recurrence were micropapillary (P = 0.021), fertility-preserving surgery (P = 0.014), and laparoscopic approach (P = 0.009). Of these 112 patients on whom lymphadenectomy was performed, 17 cases showed lymph node positive, whereas the remaining 95 patients did not. Significant differences in terms of lymph node numbers (P < 0.0001), invasive implant (P = 0.022), and International Federation of Gynecology and Obstetrics staging (P < 0.0001) were observed between the 2 groups of lymphatic node involved or not. Kaplan-Meier curves of 5-year disease-free survival revealed that there were no significant differences either between groups of lymphatic node involved or not (P = 0.778) and groups of removed nodes whether more than 10 or not (P = 0.549). CONCLUSIONS: Micropapillary, fertility-preserving, and laparoscopic approach were factors significantly affecting the recurrence of SBOT by both univariate and multivariate analysis. Lymph node metastasis did not seem to be correlated to a worse prognosis of SBOT.

Inhibition of Notch 1 receptor influenced the differentiation of Lin-CD45RA-dendritic cell precursors within ovarian carcinoma microenvironment.

BACKGROUND: Previous evidence suggested that the differentiation of Lin-CD45RA-DC precursors were prior to plasmcytoid dendritic cells (pDCs) than myeloid dendritic cells (mDCs) within ovarian cancer microenvironment. However, the mechanism is still unclear. Therefore, we investigated the function of Notch 1 signal pathway in the differentiation of Lin-CD45RA-DC precursors. METHODS: The CD34+ hematopoietic stem cells were extracted from umbilical cord blood in term parturition, and Lin-CD45RA-DC precusors were separated and induced mature. Expression of Notch1 receptor and ligands in Lin-CD45RA-DC precusors was detected by Real-time PCR and was down-regulated by shRNA or γ-secretase inhibitor (GSI). Flow cytometry was used to analyze the subset of DCs with or without SKOV3 culture supernatants. IL-12 level was detected by ELISA. RESULTS: Expression of Notch1 receptors and ligands were detected in Lin-CD45RA-DC precursor cells. The Notch1 mRNA in Lin-CD45RA-DC precursors can be down-regulated by shRNA-Notch1 lentivirus transfection and GSI. ShRNA mediated Notch 1 knock-down significantly differentiated less plasmcytoid dendritic cells (pDCs), but generated more myeloid dendritic cells (mDCs), and this would not be influenced by the supernatant of the ovarian carcinoma cell line. GSI had the same effect in the differentiation of pDC. The secretion of IL-12 significantly increased after Notch1 knock-down with or without SKOV3 culture supernatants. CONCLUSIONS: Notch1 is an important signaling pathway in the differentiation of Lin-CD45RA-DC precursor cells to plasmcytoid dendritic cells (pDCs). And this would not be affected by the supernatant of the ovarian carcinoma cell line.

Long-term outcome of patients with persistent low-level elevation of human chorionic gonadotrophin.

AIM: The aim of this study was to investigate a series of patients with sustained low-level elevated human chorionic gonadotrophin (hCG) and explore the management of these patients. METHODS: A total of 47 patients with persistent low levels of hCG were selected for analysis between January 2002 and January 2014 at the Women's Hospital of Zhejiang University, Hangzhou, China. Data were retrospectively reviewed for patient characteristics, therapeutic strategies, and follow-ups. We compared the characteristics of patients who were and were not eventually considered to have malignancies. RESULTS: Among the 47 patients, 17 with persistent low-level elevated hCG and no detectable lesions were considered to have no active malignancy. Fifteen of the 17 patients had hCG levels that returned to normal range by the end of follow-up, while the remaining two did not. The other 30 patients were eventually diagnosed as having active malignancies due to detected lesions or increasing elevation of hCG. A large proportion of these patients were diagnosed with placental site trophoblastic tumor or epithelioid trophoblastic tumor. CONCLUSION: For patients with persistent low-level elevated hCG, frequent follow-up rather than any therapy is recommended. Treatment was considered effective and safe once diagnosis of active malignancy was confirmed.

[Reliability and Validity of the Jefferson Scale of Physician Empathy in Chinese Medical Students].

OBJECTIVE: To access the reliability and validity of the student version of Jefferson Scale of Physician Empathy (JSPE-S). METHODS: The JSPE-S was translated into Chinese using back-translation procedures and administered to 358 Chinese medical students at Sichuan University. The reliability was evaluated with Split-half reliability and internal consistency. The validity was analyzed using discriminate validity, convergent validity and structure validity. RESULTS: The JSPE-S had a split-half reliability coefficient of 0.853 and Cronbach alpha of 0.861. The convergent test achieved 95.0% success rate. The discriminant test achieved 95.0% success rate. Three factors were extracted, with a cumulative variance contribution of 50.87%. The estimated factor loading ranged from 0.485 to 0.834, with factor variance ranging from 1.736 to 4.625. CONCLUSION: The Chinese JSPE-S has satisfactory reliability and validity in medical students.

Endometrial hepatoid adenocarcinoma: a rare cause of elevated serum α-fetoprotein.

Hepatoid adenocarcinoma is a rare and unusual tumor in the female genital tract. Hepatoid adenocarcinoma resembles hepatocellular carcinoma morphologically but develops in extrahepatic organs, and usually demonstrates foci of adenocarcinoma of the primary organ. Tumor cells often stain positive for anti-α-fetoprotein antibody, and may be associated with elevated serum α-fetoprotein, which may be useful as a tumor marker to guide treatment. There is little reliable information to guide clinical management of these unusual tumors and prognosis is poor despite multi-modal treatment. This report describes the diagnosis and treatment of this tumor in a postmenopausal woman.

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.

[Expression and clinical significance of GRIM-19 in epithelial ovarian carcinoma].

OBJECTIVE: To investigate the expression and clinicopathological features of gene associated with retinoid-interferon mortality-19 (GRIM-19) in epithelial ovarian carcinoma. METHODS: The expression of GRIM-19 gene in tissues from 138 cases of epithelial ovarian carcinoma, 102 cases of benign ovarian epithelial tumor and 46 cases of normal ovarian tissues were detected by Immunohistochemistry and Western blot methods. Assembled clinical survival data were analyzed with Kaplan-Meier and Cox regression models. RESULTS: The expression level of GRIM-19 in epithelial ovarian carcinoma (3.4 ± 2.0) was lower than that in benign ovarian tumor tissues (4.7 ± 2.9) and that in normal ovarian tissues (7.5 ± 2.2; P < 0.01). The level of GRIM-19 expression was related to the survival time of epithelial ovarian carcinoma patients by Kaplan-Meier analysis (P = 0.002). The shorter survival time of epithelial ovarian carcinoma patients was significantly associated with the level of GRIM-19 expression (P = 0.001), clinical stage (P = 0.001), volume of ascites (P = 0.023) and the largest diameter of the primary tumor lesion (P = 0.044) by Cox regression models. CONCLUSIONS: The low expression of GRIM-19 in the epithelial ovarian carcinoma suggests that GRIM-19 may be a key gene involved in its carcinogenesis. The expression level of GRIM-19 may be also an independent prognostic factor for epithelial ovarian carcinoma patients.

NDR1 activates CD47 transcription by increasing protein stability and nuclear location of ASCL1 to enhance cancer stem cell properties and evasion of phagocytosis in small cell lung cancer.

Small cell lung cancer (SCLC) is one of the most malignant types of lung cancer. Cancer stem cell (CSC) and tumor immune evasion are critical for the development of SCLC. We previously reported that NDR1 enhances breast CSC properties. NDR1 might also have a role in the regulation of immune responses. In the current study, we explore the function of NDR1 in the control of CSC properties and evasion of phagocytosis in SCLC. We find that NDR1 enhances the enrichment of the ALDEFLUORhigh and CD133high population, and promotes sphere formation in SCLC cells. Additionally, NDR1 upregulates CD47 expression to enhance evasion of phagocytosis in SCLC. Furthermore, the effects of NDR1 enhanced CD47 expression and evasion of phagocytosis are more prominent in CSC than in non-CSC. Importantly, NDR1 promotes ASCL1 expression to enhance NDR1-promoted CSC properties and evasion of phagocytosis in SCLC cells. Mechanically, NDR1 enhances protein stability and the nuclear location of ASCL1 to activate the transcription of CD47 in SCLC. Finally, CD47-blocking antibody can be used to target NDR1 enhanced CSC properties and evasion of phagocytosis by suppressing EGFR activation in SCLC. In summary, our data indicate that NDR1 could be a critical factor for modulating CSC properties and phagocytosis in SCLC.

Gastric-type Mucinous Carcinoma with an Abnormal Increase of CA199: A Case Report and Literature Review.

Objective: Gastric-type mucinous carcinoma (GAS), as a rare subtype of mucinous adenocarcinoma, accounts for approximately 1%-3% of cervical adenocarcinoma. It was considered as a new type of cervical mucinous adenocarcinoma by the World Health Organization (WHO) in 2014. GAS represents more aggressive disease than does usual type endocervical adenocarcinoma (UEA). Case report: A case of cervical adenocarcinoma with an abnormal increase of CA199 in a 50-year-old Chinese woman was reported. Our patient presented with abnormal vaginal discharge and combined with elevated Ca199 at the value of 2,729 U/mL. Imaging examinations showed no abnormalities. Diagnostic conical resection suggested cervical adenocarcinoma in situ. Post-operative pathology confirmed mucinous cervical adenocarcinoma (considering gastric type), infiltrating cervical interstitial >2/3, involving the deep myometrium, accompanied by vascular carcinoma infiltration and lymph node metastasis.The patients received an extensive hysterectomy and post-operative adjuvant chemoradiotherapy. The chemotherapy regimen was paclitaxel, combined with platinum. After 20 months of follow-up, the patient showed no signs of recurrence. Conclusion: Preoperative diagnosis of cervical adenocarcinoma is insidious and can be easily misdiagnosed. For patients with high preoperative Ca199, the possibility of GAS should be kept open.

[Expression and significance of LMP2 and PPM1A in gestational trophoblastic disease].

OBJECTIVE: To investigate the expression of low molecular mass polypeptide-2 (LMP2) and protein phosphatase 1A (PPM1A) in gestational trophoblastic disease and elucidate their predictive value in malignant transformation of hydatidiform mole. METHODS: The expressions of LMP2 and PPM1A protein in 196 complete hydatidiform moles (in which 28 cases with malignant transformation), 7 invasive moles, 5 choriocarcinomas and 20 normal chorionic villus were detected with the method of EnVision immunohistochemistry. Their clinicopathologic data were retrospectively analyzed. RESULTS: LMP2 and PPM1A protein expressed in cytotrophocytes, syncytiotrophoblast and extravillous trophoblast. The level of LMP2 expression in deteriorative hydatidiform mole was significantly higher than that in non-deteriorative hydatidiform mole or normal chorionic villus (6.79±2.38, 5.26±2.63 and 3.10±1.65, all P<0.01), while there were no difference compared with gestational trophoblastic neoplasms (6.42±2.68, P=0.113). The level of PPM1A expression was highest in normal chorionic villus, and decreased gradually in hydatidiform mole (non-deteriorative and deteriorative) and gestational trophoblastic neoplasms (6.30±2.98, 4.93±2.50, 4.43±2.04 and 3.33±2.06, all P<0.01); the level of PPM1A expression in deteriorative hydatidiform mole was significantly lower than that in non-deteriorative hydatidiform mole (P=0.001). The expression of LMP2 protein was correlated to theca lutein ovarian cyst, the expression of PPM1A protein was related with uterine size (P<0.05). While, there was no correlation between the expressions of the two proteins (P>0.05). CONCLUSIONS: High expression of LMP2 and low expression of PPM1A might play an important role in the motility and invasiveness of trophoblast cells and malignant transformation of hydatidiform mole. Testing the expression of LMP2 and PPM1A in hydatidiform mole tissues of initial uterine evacuation might be have some reference significance in judging outcomes of hydatidiform mole.

[Value of serum CA(125) detection for predicting recurrence and prognosis in ovarian epithelial cancer].

OBJECTIVE: To analyze the association of serum CA(125) level at the different phases with recurrence and survival, for providing simple and efficient methods about predicting recurrence and prognosis in epithelial ovarian cancer. METHODS: The clinical-pathological data from 151 patients were collected, who were histologically confirmed as primary ovarian cancer between Jan 2002 and Dec 2005. All the patients were followed up. The relationship between serum CA(125) level at different phases and clinical-pathological data were analyzed, including prognostic associated factors, 2-year or 5-year recurrent rate, 5-year survival rate, progression-free survival times, and overall survival times. RESULTS: Serum CA(125) level at pre-surgery and the end of 3-course chemotherapy were associated with most of the clinical-pathological parameters, included stage, pathological grade, amount of ascites, residual tumor size, type of recurrence, 2-year and 5-year recurrent rate, and 5-year survival rate (all P < 0.05). Progression-free survival and overall survival times were shorter in the patients with higher CA(125) level at pre-surgery or abnormal CA(125) level at the end of 3-course chemotherapy (P < 0.01). There was no relationship between the ratio of CA(125) level at pre- and post-surgery and recurrence or prognosis (all P > 0.05). CONCLUSION: Serum CA(125) level at pre-surgery and the end of 3-course chemotherapy can be used for predicting the recurrence and prognosis of epithelial ovarian cancer.

Epithelioid trophoblastic tumor that requires fertility preservation: A case report and review of literature.

OBJECTIVE: Epithelioid trophoblastic tumor (ETT) is very rare and few cases have been published in the English literature. Hysterectomy is the recommended treatment, due to the high rate of recurrence and mortality. The objective of this article is to present a rare case of ETT with fertility-preserving treatment and review published similar cases. CASE REPORT: We report the case of ETT in a 19-year-old Chinese woman, who had a strong desire of fertility preservation. She presented with vaginal spotting and hysteroscopy showed an isolated solid mass (2.0 × 1.5 × 1.5 cm) at the right corner of the uterine cavity. Serum ß-human chorionic gonadotropin (ß-HCG) persisted at low level elevation about 100 IU/L. We treated her with a lesionectomy and 3 cycles EP-EMA (etoposide, cisplatin/etoposide, methotrexate and actinomycin) chemotherapy regimen. The patient is now in stable condition, without any signs of recurrence during 20 months of follow-up. CONCLUSION: Fertility-preserving surgery would probably be a feasible and safe strategy for the patients whose lesions can be completely removed.

[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.

Synchronous occurrence of benign mesothelioma and adenomatoid tumor of uterus: A case report and review of literature.

INTRODUCTION: Synchronous occurrence of benign cystic mesothelioma and adenomatoid tumor of uterus (UAT) are very rare and few cases have been published in the English literature. They are easily misdiagnosed as malignant by clinicians, due to the lack of reports. PATIENT CONCERNS: A case of benign mesothelial combined with uterus adenomatoid tumor (UAT) in a 48-year-old Chinese woman was reported. Our patient presented with abdominal pain and surgery showed a large subserous mass (12.0 × 11.4 × 9.8 cm) combined with a small intramural solid nodule (2.0 × 1.0 × 1.0 cm), and multiple minute neoplastic growth on the ovary. DIAGNOSIS: Due to the patient's symptoms, pathological findings, she was diagnosed with synchronous occurrence of benign mesothelioma and UAT. INTERVENTIONS: We treated her with a total hysterectomy and bilateral adnexectomy. OUTCOMES: The patient is now in stable condition, without any signs of recurrence during 1 year of follow-up. LESSONS: Most mesotheliomas are malignant, synchronous occurrence of benign mesothelioma and UAT are extremely rare. And they are often misdiagnosed as malignancy by clinicians. Our case report can improve the awareness of the disease and avoid excessive treatment.

[Case report and literature review of epithelioid trophoblastic tumor].

OBJECTIVE: To evaluate the clinical characteristics of epithelioid trophoblastic tumor (ETT). METHODS: Six cases of ETT treated in Women's Hospital, School of Medicine, Zhejiang University from 2005 to 2007 were retrospectively analyzed, together with a literature review. RESULTS: Six cases of ETT were diagnosed pathologically after surgery. The age of patients ranged from 27 to 46 years. The most common presentation was abnormal vaginal bleeding (5/6). The preceding gestational events were hydatidiform mole in 1 case, abortion in 2 cases, and term delivery in 3 cases. The interval between the preceding gestation and the diagnosis of ETT ranged from 15-48 months. The serum human chorionic gonadotropin (hCG) level was 46-121.47 IU/L. Four cases presented with metastasis, including lung metastasis in all of the 4 cases, liver metastasis in 1 case, and pancreas metastasis in another 1 case. The main therapies were surgery combined with chemotherapy. All of the 6 cases received total abdominal hysterectomy, and 1 case also had lung lobectomy. One case had a recurrence but refused any treatment again, and was lost to follow up; the therapy of 1 case unfinished; another 4 cases were without evidence of disease 9 to 19 months after surgery. CONCLUSIONS: The confirmation of ETT diagnosis is difficult before surgery. Surgical management is mostly recommended in ETT. The role of chemotherapy in ETT is not clear yet.

[Study on parental origin of complete hydatidiform mole detected by polymorphisms of microsatellite].

OBJECTIVE: Through detection of 9 loci of polymorphisms of microsatellite to investigate the parental origin of complete hydatidiform mole. METHODS: Using the technology and method of multipolymerase chain reaction and electrophoresis in denatured polyacrylamide gel and silver stain detection, we carried out DNA analysis on 50 cases of complete hydatidiform mole diagnosed by histopathology and 50 copies of the couples' peripheral blood. RESULTS: There are 7 cases of biparental complete hydatidiform mole (14%); and 43 cases of androgenetic complete hydatidiform mole (86%) among 50 cases of complete hydatidiform mole. CONCLUSIONS: Detection of polymorphisms of microsatellite is a technology that can be used to identify the parental origin of complete hydatidiform mole. It is simple, quick, reliable and highly efficient.

[Sensitivity of PTEN gene-transfected endometrial carcinoma cell line to doxorubicin-induced apoptosis].

OBJECTIVE: To investigate whether PTEN can increase sensitivity of Ishikawa cells, an endometrial carcinoma cell line, to doxorubicin. METHODS: Ishikawa cells transfected by PTEN gene or not were separately treated with serial concentrations of doxorubicin. The sensitivity of cells to doxorubicin was determined by MTT assay. The cells were stained with Hoechst 33258 and examined under fluorescence microscope to determine cell apoptosis. Immunoprecipitation and Western blotting analysis were performed to evaluate the effects of doxorubicin on phosphorylation of Bad and Akt/PKB. RESULTS: Doxorubicin induced cell death of the PTEN-transfected and non-transfected Ishikawa cells in a dose-dependent manner, but the cell death was more significant in PTEN-expressing clones than in parental Ishikawa cells. A low concentration of doxorubicin (0.1 micromol/L) did not affect cell apoptosis in PTEN-null Ishikawa cells, but it induced cell apoptosis in PTEN-expressing clones. A high concentration of doxorubicin (1 micromol/L) induced cell apoptosis in both cell lines. However, the percentage of apoptotic cells was higher in PTEN-expressing clones than that in parental Ishikawa cells. In the PTEN-expressing clones, expression of phospho-Akt/PKB and phospho-Bad (Ser-136) was down regulated. Doxorubicin reduced the levels of phospho-Akt/PKB and phospho-Bad (Ser-136) in both cell lines, but the most significant reduction occurred in the PTEN-expressing clones. CONCLUSION: PTEN significantly enhances chemosensitivity of Ishikawa cells to doxorubicin. With PTEN expression, doxorubicin may exert apoptosis-induction activity by downregulation of the PI3k/Akt/PKB signaling pathway in Ishikawa cells.

[Inhibitory effect of tumor suppressor PTEN on cell growth of endometrial carcinoma].

OBJECTIVE: To evaluate the inhibitory effect of tumor suppressor PTEN on cell growth of endometrial carcinoma. METHODS: The exogenous wild PTEN cDNA via an adenoviral vector (Ad-PTEN) was introduced into Ishikawa cells. The expression of PTEN protein was detected by Western blot. The growth of Ishikawa cells was evaluated by trypan blue exclusion method and MTT. RESULTS: The expression of PTEN protein was induced on day 1, and greatly increasing on day 3 - 5 after Ad-PTEN infection. The expression of PTEN significantly inhibited the growth of Ishikawa cells, and also significantly inhibited the growth of Ishikawa cells induced by IGF-II. CONCLUSION: Adenovirus-mediated introduction of exogenous PTEN into human endometrial carcinoma cells can induce growth suppression. PTEN gene may be a novel therapeutic agent for endometrial carcinoma.