Safety and Efficacy of Neoadjuvant Chemotherapy Followed by Radical Surgery Versus Radical Surgery Alone in Locally Advanced Cervical Cancer Patients.

OBJECTIVES: This study aimed to evaluate the safety and efficacy of neoadjuvant chemotherapy (NACT) followed by radical surgery (RS) among patients with locally advanced cervical cancer (LACC). METHODS: Eight hundred patients with LACC received either NACT followed by RS (NACT-RS) or RS alone. The primary outcome measures assessed the efficacy and adverse effects of NACT. Secondary outcome measures compared the preoperative clinical stage to the postoperative pathologic stage in NACT-RS and RS patients, assessed intraoperative and postoperative complications, including the adverse effects of postoperative radiotherapy and radiochemotherapy, and estimated the 5-year progression-free survival and 5-year overall survival. RESULTS: The clinical response to NACT was 89.54%. Patients in the NACT-RS group had lower preoperative hemoglobin levels (115.20 vs 122.04 g/L, P < 0.001), a longer operative time (mean, 233.66 vs 224.37 minutes, P = 0.008), more intraoperative bleeding (750.34 vs 684.41 mL, P = 0.011), a shorter duration of catheter use (mean, 29.84 vs 32.14 days, P = 0.036), and a lower incidence of postoperative complications (7.30% vs 13.62%, P = 0.002) and postoperative radiotherapeutic and radiochemotherapeutic adverse effects (3.16% vs 4.63%, P < 0.001) compared to patients in the RS group. The 5-year progression-free survival and 5-year overall survival were 80.30% and 81.10% in the NACT-RS group and 81.00% and 78.50% in the RS group (P > 0.05). Pathological poor differentiation, nonsquamous cell carcinoma, parametrial invasion, positive pelvic lymph node, and lymphovascular invasion (P < 0.05) were independent risk factors for recurrence. CONCLUSIONS: Neoadjuvant chemotherapy may reduce RS-associated complications and postoperative radiotherapeutic and radiochemotherapeutic adverse effects in Chinese patients with LACC.

[The regulation mechanism of MASPIN expression in ovarian carcinoma].

OBJECTIVE: To study the regulation of P63 on expression of MASPIN in ovarian cancer by observe MASPIN promoter activity changes before and after transient transfection of constructed P63 and MASPIN reporter gene plasmids. METHODS: The MASPIN reporter plasmid, fused with luciferase reporter gene, was constructed and transfected into SKOV3 cells together with P63 (TAP63, ANP63) express plasmid transiently. The MASPIN promoter activity was determined in both the transfected cells and controlled ones by Luciferase Assays and the transcription of MASPIN mRNA of them was evaluated with semi quantitative RT-PCR. RESULTS: The MASPIN reporter plasmid was successfully constructed and transiently transfected into SKOV3 cells together with P63 (TAP63, ANP63) expression plasmid. The data showed among the tested P63 splice variants, TAP63 remarkably activated MASPIN promoter transactivation (P < 0.05). No significant difference in the activity level of MASPIN promoter was detected in the SKOV3-vector and SKOV3-ANP63 cells (P > 0.05). The level of MASPIN mRNA expression was notably enhanced in SKOV3-TAP63 cell after transient transfected with TAP63 express plasmid (P < 0.05), but no significant difference among the SKOV3, SKOV3-vector and SKOV3-ANP63 cell (P > 0.05) was detected. CONCLUSION: TAP63 can activate the transcription activity of MASPIN promoter, as well as regulate the expression of MASPIN. Put all together, these results suggested that MASPIN is a new molecular target of P63.

Involvement of ARLTS1 in chemotherapy and apoptosis in ovarian cancer cell line.

PURPOSE: Recent evidence suggests that ADP-ribosylation factor-like tumor suppressor gene 1(ARLTS1) may act as a tumor suppressor gene. However, its role in tumor chemotherapy remains unclear. The aim of this study is to investigate the effects of ARLTS1 gene in regulation of chemosensitivity in ovarian cystadenocarcinoma cell line SKOV3. METHODS: We stably expressed wild-type (wt) ARLTS1 and empty vector (neo) in SKOV3 cells. Chemosensitivity test was carried out with four chemotherapeutic agents. Cell proliferation, cycle kinetics and apoptosis were evaluated by MTT assay and flow cytometry. Apoptosis-related proteins caspase-3 and bcl-2 were determined by Western blot analysis. RESULTS: The proliferation of wtARLTS1 clones was more dramatically inhibited by all the cytotoxic agents than parental cells (P < 0.05). Increased sensitivity to chemotherapy by two to threefolds was detected in wtARLTS1 cells. The rate of apoptosis in wtARLTS1 was 60.2% treated with DDP (10× peak plasma concentration, PPC), which was dramatically higher than that of neo and parental cells (P is 0.017 and 0.020, respectively). Expression of caspase-3 and bcl-2 in parental cells declined modestly when treated with DDP, while in wtARLTS1 clones the expression of caspase-3 and bcl-2 levels declined more dramatically and become undetectable at lower DDP doses (P = 0.023 and <0.001, respectively). CONCLUSION: Our findings suggested that ARLTS1 may facilitate chemosensitivity in ovarian cancer cells by acting synergistic with chemotherapeutic agents to induce the apoptosis signaling pathway and regulate apoptosis-related proteins.

[Effect of autophagy gene Beclin 1 on the growth of cervical cancer HeLa cells in vitro and vivo].

OBJECTIVE: To investigate the effects of autophagy gene Beclin 1 on growth of cervical cancer HeLa cells in vitro and vivo. METHODS: The eukaryotic expression vector of Beclin1 was constructed and transfected via lipofectamine into HeLa cells. The experimental cells were classified into 3 groups: pcDNA3.1(+)-Beclin1 group,pcDNA3.1(+) group and HeLa group. Real time-ploymerase chain reaction and Western blot were used for detecting expression of Beclin1 mRNA and protein in the transfected cells. Flow cytometry (FCM) was employed to observe the effect of transfection on the apoptosis of HeLa cells, and proliferation was analyzed by MTT assay. The formation of autophagic vacuoles was measured by MDC staining. HeLa cells transfected with plasmid pcDNA3.1(+)-Beclin1 and pcDNA3.1(+) were inoculated subcutaneously in nude mice. The carcinogenic and growth activities of cancer cells in vivo were observed. RESULTS: Eukaryotic expression vector pcDNA3.1(+)-Beclin1 was constructed successfully. It significantly improved the expression of Beclin1 mRNA and protein in HeLa cells. The proliferation of HeLa cells was inhibited, and the inhibition rate was 58.7%. FCM investigation showed that the apoptotic rate was (28.22 ± 2.34)% of pcDNA3.1(+)-Beclin1 group, significantly higher than the (14.6 ± 4.6)% in the pcDNA3.1(+) group and (11.2 ± 3.0)% in the HeLa group (P < 0.05). The monodansylcadaverin (MDC) staining showed significantly more autophagic vacuoles in the pcDNA3.1(+)-Beclin1 group (10.9%) than that in the pcDNA3.1(+) group (3.1%) and HeLa group (2.5%) (P < 0.05). After transfected with vector pcDNA3.1(+)-Beclin1, the carcinogenic activity of HeLa cells was decreased in nude mice, and the inhibition rate of tumor growth was 52.2%. CONCLUSIONS: Autophagy gene Beclin 1 overexpression can inhibit the proliferation and growth of HeLa cells in vitro and vivo,while promote autophagy and apoptosis of HeLa cells. So it might be one of new gene therapy strategies for cervical carcinoma.

Beclin1 induces autophagy and its potential contributions to sensitizes SiHa cells to carboplatin therapy.

BACKGROUND: To investigate the role of Beclin1 in the relationship between autophagy and apoptosis after carboplatin treatment. METHODS: After overexpression or partial silencing of Beclin1 in cervical cancer SiHa cells, the transfected group and the control group were treated with carboplatin for 48 hours. The expression of Beclin1 and LC3 was measured using a western blot. The ultrastructural analysis was conducted with an electron microscope. Fluorescent changes in autophagic cells were observed using reverse fluorescent microscopy. The percentage of apoptotic cells and autophagic cells and cell proliferation were assessed by flow cytometry and MTT assay. RESULTS: Expression of LC3 and Beclin1 protein was up-regulated in overexpressed transfectants of SiHa cells. After treatment with carboplatin for 48 hours, the flow cytometric analysis indicated that the Beclin1 transfected group showed a greater increase in apoptosis and autophagic cells than did the nontransfected group. MTT assay showed that carboplatin inhibited proliferation of SiHa cells in Beclin1-overexpressed transfectant cells more than in nontransfectant cells. CONCLUSIONS: These results suggest that autophagy and apoptosis had differential contributions to carboplatin-induced death of cervical cancer SiHa cells. Overexpression of Beclin1 in SiHa cells may enhance apoptosis signaling induced by carboplatin.

Association of the ARLTS1 variants with familial ovarian cancer risk in China.

ARLTS1 has been identified in chromosome 13q14 as a tumor suppressor gene of the adenosine diphosphate-ribosylation factor family with pro-apoptotic characteristics. The ARLTS1 mutation Trp149Stop and Cys148Arg have been shown to be associated with familial cancers, but limited information is available regarding the impact of ARLTS1 variants on familial ovarian cancer (OC). The aim of this study was to evaluate the ARLTS1 genetic variants associated with familial OC risk in China. We genotyped 85 OC patients with family ovarian/breast history, 80 sporadic OC patients, and 120 controls from general population by denaturing high-performance liquid chromatography screening analysis followed by direct sequencing of the conspicuous polymerase chain reaction products. ARLTS1 Cys148Arg revealed a significant association with an increased risk of familial OC compared with both sporadic cases and controls in a dose-dependent manner (P = 0.0031 and 0.012, respectively). In the clinical-pathological study, our results support previous data in demonstrating that familial OC was associated with younger age at diagnosis (49.7 years vs 53.3 years; P = 0.014), higher proportion of tumors of advanced stages (81.2% vs 67.5%; P = 0.033), and higher rates of serous adenocarcinomas (76.4% vs 53.8%; P = 0.028) compared with sporadic OC cases. To investigate the association between genetic variants of ARLTS1 and the clinical-pathological characteristics of familial OC, we identified a significantly higher proportion of serous adenocarcinoma (55/67, 82.1%) and higher rates of advanced stage tumors (88.1% vs 55.6%; P = 0.004) in ARLTS1 Cys148Arg carriers. We showed a significantly increased risk of familial OC for ARLTS1 Cys148Arg variant, which indicate that ARLTS1 may play a role in familial OC.

[Expression of THY1 gene in epithelial ovarian cancer].

OBJECTIVE: To detect the expession of THY1 in ovarian serous cystadenocarcinoma tissues. METHODS: Immunohistochemistry was performed to detect the expression of THY1 gene in formalin-fixed, paraffin-embedded specimens of normal ovaries (n = 25), ovarian serous cystadenoma (n = 25), and serous cystadenocarcinoma (n = 53). The correlation of THY1 expression with clinicopathological parameters was statistically analyzed. RESULTS: The positive expression rates of THY1 protein in normal ovaries, ovarian serous cystadenomas and ovarian serous cystadenocarcinomas were 60.0% (15/25), 72.0% (18/25) and 34.0% (18/53), respectively. The values of IOD of THY1 protein expression were 288,449.2 +/- 60,087.3, 271,655.6 +/- 66,588.7 and 252,087.6 +/- 45,559.4, respectively. The expression of THY1 protein was significantly down-regulated in ovarian serous cystadenocarcinoma tissues compared with that in normal ovarian tissues and ovarian serous cystadenoma tissues (P < 0.05). THY1 expression was negatively correlated with surgical-pathological staging, histological differentiation and lymph node involvement (P < 0.05). CONCLUSION: The decreased level of THY1 expression may be related with the occurrence and development of ovarian serous cystadenocarcinoma.

[Clinicopathologic features of 234 cases with borderline ovarian tumors].

OBJECTIVE: To determine the clinicopathologic characteristics and prognostic factors that may be used to predict the poor outcome of patients with borderline ovarian tumors. METHODS: All cases with borderline ovarian tumors treated in the West China Second University Hospital from January 2001 to June 2007 were analyzed retrospectively for clinicopathologic features, treatment parameters and outcome of treatment. Univariate and multivariate analyses were used to assess independent prognostic factors using the logistic regression model. RESULTS: The median age of 234 patients was 40.1 years with a range of 14 to 80 years. There were 101 (43.2%), 94 (40.2%), 19 (8.1%), 12 (5.1%), 8 (3.4%) cases of serous, mucinous, mixed, endometrioid and clear cell tumors, respectively. Out of 234 cases, 182 (77.8%) underwent laparotomy and 45 (19.2%) underwent laparoscopy. Seven women underwent laparoconversion. Fertility sparing surgery was performed on 119 cases (50.9%) and radical surgery was performed on 115 cases (49.1%). Totally 161 (68.8%) patients had stage I, 19 (8.1%) had stage II, 54 (23.1%) had stage III, and none had stage IV disease. Sixty-four women received postoperative chemotherapy. The median follow-up was 40 months with a range of 8 to 78 months. Recurrence was found in 26 cases (11.1%) during follow-up, and no tumor-related death was reported. The logistic regression model showed that surgery procedure (OR = 2.304, P = 0.024), cyst rupture (OR = 2.213, P = 0.038), stage (OR = 4.114, P < 0.01), microinvasion (OR = 2.291, P = 0.046) and peritoneal implants (OR = 2.101, P = 0.016) were the five independent prognostic factors affecting recurrence. CONCLUSIONS: Although patients with borderline ovarian tumors have an excellent prognosis, the risk of recurrence remains in some patients. Emphasis should be put on these patients with high risk factors and preventive strategies should be taken to prevent their progression.

[Diagnosis and management of 57 cases with rectovaginal endometriosis].

OBJECTIVE: To study the diagnosis and therapy of the rectovaginal endometriosis. METHODS: Clinical data of 57 women with rectovaginal endometriosis admitted to the West China Second University Hospital of Sichuan University in last two years,were retrospectively reviewed. RESULT: The average age of patients was 40.1 years. The main clinical manifestations were dysmenorrheal, changes of menorrhea and digestive stimulation. The diameter of deep endometriosis nodules was between 1-6 cm, and 77% were found to have more than one nodules. Seven of these patients had positive results in transvaginal ultrasonography; 61%(11/18) patients had elevated CA125 levels. Thirteen patients were given preoperational medical treatment, but had no effect. All patients, except one accepted laparotomic therapy of complete excision of endometriosis nodules; 23 cases underwent drug therapy after operation. No patients had recurrence in recto-vaginal septum after complete excision; only one recurred in right ovary. Patients who failed to remove the total lesion showed improvement in pain. CONCLUSION: Diagnosis of the rectovaginal endometriosis is based on symptoms, vaginal and rectal examination, and auxiliary examination. Complete excision of endometriosis nodules is the main therapeutic method.

[Expression of maspin and its relation to tumor vascularization in epithelian ovarian cancer].

OBJECTIVE: To investigate the correlation of Maspin expression with angiogenesis in ovarian cancer. METHODS: The expression of Maspin, VEGF, heparanase gene was detected by immunohistochemistry in formalin-fixed, paraffin-embedded specimens of normal ovaries (n=31) and epithelial ovarian cancers (n=35). Correlations of Maspin, VEGF and heparanase expression with clinicopathological parameters of ovarian tumors were statistically analyzed. Microvessel density (MVD) was counted by endothelial cells immunostained with anti-CD34 monoclonal antibody. RESULTS: The expression of Maspin, VEGF, heparanase protein were up-regulated in ovarian cancer cells compared with control group (P < 0.05). Significant association was observed in Maspin protein expression and clinical parameters such as non-serous ovarian carcinoma, lower tumor stage and non-ascites (P < 0. 05). VEGF protein expression correlated with ascites positively (P < 0.05). Heparanase protein expression was identified obviously to correlate with lymph node involvement and ascites (P < 0.05). MVD has significant correlation with lymph node involvement and tumor stage (P < 0.05). Maspin protein expression correlated negatively with VEGF and MVD(P < 0.05), but had no relationship with heparanase (P > 0.05). Significant direct association was demonstrated between VEGF protein expression and MVD (P < 0.05), but VEGF protein expression had no correlation with heparanase. CONCLUSION: Maspin might have inhibitory effects on invasion and metastasis of epithelian ovarian cancer.

[Polymorphism of ARLTS1 gene in early-onset ovarian cancer].

OBJECTIVE: To evaluate the impact of ARLTS1 gene mutations among Chinese Han-nationality patients with early-onset epithelial ovarian carcinoma (OC) in Southern China. METHODS: Tumor and blood samples collected from early-onset, late-onset epithelial ovarian carcinoma patients and control group were collected. After purification of DNA, the ARLTS1 gene fragments were amplified with PCR. The DHPLC was applied to analyze the PCR products. The mutation products were confirmed with DNA sequencing analysis. RESULTS: ARLTS1 Cys148Arg revealed a significant association with an increased risk in early-onset OC group compared with both late-onset cases and controls. ARLTS1 Ser99Ser revealed no impact on both early-onset OC group and late-onset cases. CONCLUSION: A significantly increased risk for young ovarian cancer patients with ARLTS1 Cys148Arg variant were observed, which indicate that ARLTS1 may play a important role in early-onset of OC.

[Effects of ARLTS1 gene on growth and apoptosis of epithelial ovarian cancer SKOV3 cells].

OBJECTIVE: To investigate the effect of ARLTS1 (ADP-ribosylation factor-like tumor suppressor gene 1) on the growth and apoptosis of ovarian cystadenocarcinoma cell line SKOV3. METHODS: The recombinant plasmid pCMV-Tag 3B-ARLTS1 was constructed and transfected into SKOV3 cells by liposome protocol. The expression of ARLTS1 mRNA and its protein were examined by RT-PCR and Western blotting. Effects of ARLTS1 gene on growth and apoptosis of the transfected cells were evaluated by MTT assay and flow cytometry (FCM). The alterations of caspase-3 and bcl-2 protein levels were examined by Western blotting. RESULTS: The stable transfection of pCMV-Tag 3B-ARLTS1 in SKOV3 cells were obtained and verified by RT-PCR and Western blotting methods. After transfected with ARLTS1, more SKOV3 cells were observed in S phase by FCM compared with those in the other two groups (P is 0.035 and 0.011, respectively). The 36.7% apoptotic index of ARLTS1 transfected cell was significantly higher than that of the other two groups (P < 0.001). The growth of ARLTS1 transfected cells was dramatically inhibited compared with cells transfected with Vector (P < 0.05). Western blotting indicated a significantly decrease in caspase-3 and bcl-2 protein levels in cells transfected with ARLTS1 compared with SKOV3 cells (P is 0.021 and 0.013, respectively). CONCLUSION: ARLTS1 transfection can inhibit proliferation and induce apoptosis of SKOV3 cells in vitro and down-regulate the expression of caspase-3 and bcl-2. ARLTS1 gene may be a candidate tumor suppressor gene in ovarian cancer.

[Proliferation and apoptosis effect of rosiglitazone on human ovarian cancer cell line SKOV3].

OBJECTIVE: To study the effect of PPARgamma, activated by Rosiglitazone (RSG), on the regulation of cell cycle and apoptosis in human ovarian cancer cell line SKOV3. METHODS: SKOV3 was treated with various levels of RSG (0.1, 1.0, 10 and 100 micromol/L), its proliferation was evaluated by MTT assay, apoptosis was determined by Hoechst33258 fluorescence and PI staining flow cytometry, and cell cycle analyzed by flow cytometry, respectively. The expression of PPARgamma, c-myc mRNA and protein were investigated by means of RT-PCR and immunocytochemistry respectively. RESULTS: MTT assay indicated that RSG could inhibit the proliferation of SKOV3 in a concentration-dependent and time-dependent pattern. PI staining flow cytometry data demonstrated that the apoptosis of SKOV3 and cell cycle was arrested at G1 stage induced by RSG in a concentration-dependent manner, after 24h inducing with RSG at the concentration of 10, 100 micromol/L, the apoptosis rates were 45.75% and 51.97% respectively, which were significantly higher than that of control group. Hoechst33258 fluorescence staining demonstrated that the apoptosis bodies were found in the SKOV3 treated with RSG. RT-PCR and immunocytochemistry indicated human ovarian cancer SKOV3 cells expressed PPARgamma. PPARgamma mRNA and protein expression increased, while c-myc mRNA and protein expression decreased in SKOV3 cells treated with RSG after 24 h. (P < 0.05). CONCLUSIONS: RSG significantly inhibits SKOV3 survival and induces its apoptosis by stoppage in G1 phase. The effect of growth inhibition is associated with downregulating expression of c-myc mRNA and protein via activation of PPARgamma.

[Expression and clinical significance of CDC6 and hMSH2 in cervical carcinoma].

OBJECTIVE: To detect the expression of CDC6 and hMSH2 in cervical carcinoma, explore the correlation with clinical significance and pelvic lymph node metastases. METHODS: SP immunohistochemistry technique was used to detect the expression of CDC6 and hMSH2 in specimens of 70 cervical carcinoma, and 14 normal cervix. Correlation between the expression of CDC6, hMSH2 and the clinicopathologic factors of cervical carcinoma were statistically analyzed. RESULTS: The rates of positive expression of CDC6 and hMSH2 were 70.0%, 54.3% in cervical carcinoma, and significantly higher than those in normal cervix (P=0.000, 0.006). The CDC6 expression rate was related to pathological subtype, FIGO stage, histological tumor grade, pelvic lymph node metastases (P<0.05). The hMSH2 expression rate was related to pathological subtype, FIGO stage and pelvic lymph node metastases (P<0.05), but did not associate with histological tumor grade. The expression of CDC6 was positively correlated with hMSH2 in cervical cancer (r(s)=0.338, P=0.004), meanwhile it was not correlated with hMSH2 in 19 patients with pelvic lymph node metastases. CONCLUSION: CDC6 and hMSH2 play an important role in the invasion and metastasis of cervical carcinoma. CDC6 and hMSH2 may be useful for predicting prognosis of cervical carcinoma.

[Expression of VEGF and its significance in primary diffuse large B-cell lymphoma of the female genital system].

OBJECTIVE: To investigate the expression of vascular endothelial growth factor (VEGF) and its significance in primary diffuse large B-cell lymphoma of the female genital system. METHODS: VEGF mRNA and expression of VEGF protein were detected by real-time fluorescence quantitative PCR method and immunohistochemistry, respecitvely. Microvessel density (MVD) was obtained after staining of microvessels using CD34 antibody. The relationship between the expression of VEGF and MVD, clinical features and prognosis of primary diffuse large B-cell lymphoma of the female genital system patients were analyzed. RESULTS: (1) Over expression of VEGF mRNA was found in 17/20 (85.0%) by real-time fluorescence quantitative PCR method and the tumor cells expressed VEGF protein in 48/56 (85.7%) by immunohistochemistry; (2) The expression of VEGF was positively correlated to the MVD of the tumor (r = 0.76, P = 0.0170); (VEGF expression was correlated to clinical stages, B symptom and the serum level of lactate dehydrogenase (LDH) (r = 0.77, P = 0.044; r = 0.58, P = 0.023; r = 0.69, P = 0.017); (4) Cox multivariate analysis demonstrated that the age of more than 60 years and the positive VEGF expression were independent prognostic factors (P < 0.05). CONCLUSION: VEGF was widely expressed in the tumor cells of primary diffuse large B-cell lymphoma of the female genital system and may be related to the clinical features and prognosis of primary diffuse large B-cell lymphoma of the female genital system patients.

[The effects of progesterone and insulin-like growth factor (IGF- I and IGF-II) on proliferation in human decidual stromal cells of early pregnancy in vitro].

OBJECTIVE: To study the effect of progesterone and Insulin-like growth factor (IGF-I and IGF-II) on proliferation in human decidual stromal cells of early pregnancy in vitro. METHODS: [3H]Thymidine (3H-TdR) uptake was applied to assess cell proliferation in human decidual stromal cells of early pregnancy (gestation of 5 to 7 weeks) in vitro after cultured with progesterone, IGF-I or IGF-II. RESULTS: Progesterone, IGF-I and IGF-II stimulated cell proliferation by 1.6-3.4 folds inhuman decidual stromal cells of early pregnancy in vitro (P<0.01), and those effects were time-dependent (P<0.01). CONCLUSION: Progesterone, IGF-I and IGF-II may play an important role in the regulation of proliferation and decidualization of stromal cells in human decidua of early pregnancy, which is essential for embryo implantation and the maintenance of early pregnancy.

[Improve the chemosensitivity of resistant ovarian cancer cell by small RNA interference].

OBJECTIVE: To investigate the effects of siRNA on the drug resistance reversal of ovarian cancer cell. METHODS: The siRNA was transfected into human ovarian cancer cell line OVCAR8/TR by liposome. ATP-bioluminence assay was applied to measure the drug sensitivity to four chemotherapeutic agents before and after transfection. RESULTS: ATP-bioluminence assay showed that OVCAR8/TR cells were resistant to cDDP, ADM and Taxol. After siRNA transfection, OVCAR8/TR cells were sensitive to ADM and Taxol which are tansported by P-gp. The inhibition rate of ADM was improved from 37% to 58%, and that of Taxol was improved from 26% to 78%. However, the resistance of OVCAR8/TR cells to cDDP was not reversed. CONCLUSION: siRNA can effectively improve the drug resistance to chemotherapeutic agents which are transfered by P-gp. The RNA interference can reverse MDR1-mediated drug resistance in ovarian cancer cell.

[Effect of siRNA inhibiting HPV16 E6 gene expression on proliferation and cell cycle of cervical cancer cell line CaSki].

OBJECTIVE: To observe whether human papillomavirus 16 (HPV16) E6-specific small interfering RNAs (siRNAs) can be employed to inhibit the growth of cervical cancer cell line, and to investigate the associated mechanism. METHODS: RNAi was performed using synthetic small interfering RNAs transferred into CaSki cell line by lipofectamine. The cell growth curves, live cell ratio and inhibition ratio of cells were measured by using cell counting. At various time points of post-transfection, the distributions of cell cycle, the expression levels of HPV16 E6, p53, p21 mRNA and proteins were detected by using flow cytometry (FCM) and real-time quantitative reverse transcription-polymerase chain reaction (real-time RT-PCR). RESULTS: The growth inhibition of E6 siRNA to CaSki cells was demonstrated after cells treated with E6 siRNA. No substantial G1 arrest was observed by FCM analysis. For 24 hours after cell transfection, the level of E6 mRNA was decreased by 20. 11 folds compared with control (P < 0.05). However, p53 and p21 mRNA levels appeared unaffected. 48 hours after cell transfection, the expression level of E6 protein was efficiently decreased, but the P53 and P21 protein levels increased in comparison. CONCLUSIONS: The inhibitory effect of HPV16 E6 siRNA to CaSki cell maybe due to specially and efficiently silence E6 mRNA expression, decrease the degradation of wild type P53 protein, and then recover the function activity of P53 protein.

[Expression of metastasis suppressor gene KAI1 in cervical carcinoma and infections of HPV16 E6, E7 and HPV18 E6/E7].

OBJECTIVE: To investigate the expression of metastasis suppressor gene KAI1 in cervical carcinoma and the impact of human papillomavirus 16 E6, E7, 18 E6/E7 infection on the expression of KAI1. METHODS: The expressions of KAI1 protein in the formalin-fixed, paraffin-embedded specimens of 20 normal cervical epthelium, 15 cervical in situ carcinoma and 70 primary invasive cervical carcinoma were detected by immunohistochemistry SP. Polymerase chain reaction (PCR) tests were also undertaken to detect the HPV16 E6, E7 and HPV18 E6/E7 DNA. RESULTS: The expression of KAI1 protein was down-regulated in the invasive carcinoma and in situ carcinoma compared with the controls (P < 0.05). No significant difference was found in the expression of KAI1 protein between invasive carcinoma and in situ carcinoma. The infections of HPV16 E6, E7 and HPV18 E6/E7 were found in 67.1%, 54.3% and 12.9% of the invasive carcinoma, respectively. However, there was no correlation between the expression of KAI1 and the infections of HPV16 E6, E7 and HPV18 E6/E7. CONCLUSION: The expression of KAI1 protein is down-regulated in cervical carcinoma, which is not associated with the infection of HPV16 E6, E7 and 18 E6/E7.

Mifepristone attenuates depression-like changes induced by chronic central administration of interleukin-1ß in rats.

Increased proinflammatory cytokines, such as interleukin (IL)-1ß, may play an important role in the etiology of depression because they cause the hypothalamic-pituitary-adrenal axis to release glucocorticoids (GC) and induce dysfunction of serotonin and norepinephrine neurotransmission. Sustained increase in GC may activate microglia to induce neuroinflammation, and suppress astrocytes to produce neurotrophins, which lead to neuronal apoptosis. Here, we tested the hypothesis that glucocorticoid receptor (GR) antagonist mifepristone (RU486) may attenuate IL-1ß-induced depression-like behavior by regulating the neuroinflammation and neurotrophin functions of microglia and astrocytes. Rats received intracerebroventricular injections of IL-1ß (10 ng) and/or subcutaneous injections of RU486 for 14 days. Then animal depression-like behaviors, serum corticosterone concentration, the levels of pro-inflammatory cytokines (TNF-α, IL-6), mRNA and protein expressions of CD11b, GFAP and neurotrophins (pro-BDNF, BDNF, GDNF and their receptors TrkB, p75, GFRα-1 and GFRα-2) in the amygdala were studied. Compared to controls, significantly decreased rearing score and increased defecation in the open field test, decreases in ratio of open/closed time in the elevated plus maze and in sucrose preference, while increased level of corticosterone in the serum were found in the rats administrated with IL-1ß. IL-1ß administration also reduced the expressions of GFAP, BDNF, GDNF and its receptor GFR-α1, but increased the expressions of CD11b, pro-BDNF, p75 and pro-inflammatory cytokines (TNF-α, IL-6) concentrations. RU486 treatment markedly attenuated these changes induced by IL-1ß, except for the expressions of GFR-α1. In conclusion, RU486 may improve depression-like changes by suppressing microglia and inflammation and promoting astrocytes to restore neurotrophin function.