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Objective@#To investigate the effects of overexpression of microRNA-7 (miR-7) on the proliferation and invasion of HepG2 cells and the underlying mechanism in vitro.@*Methods@#The relative expression levels of miR-7 and Raf1 in hepatocellular carcinoma (HCC) tissues and adjacent normal tissues (ANT) were detected by quantitative real time-PCR (qRT-PCR). The relationship between the expression of miR-7 and the characteristics of HCC patients was analyzed. Cells were divided into blank control group, negative control (NC) group and miR-7 mimics transfected group, miR-7 mimics and NC were transfected into HepG2 cells by Lipofectamine™2000. The relative expression of miR-7 was detected by qRT-PCR. The proliferation ability of HepG2 cells was detected by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay. The invasion of HepG2 cells was detected by Transwell assay. The target genes of miR-7 were predicted by TargetScan and the binding effect of miR-7 on the 3′UTR of Raf1 was verified by dual luciferase reporter assay.The expressions of Raf1 protein in hepatocellular carcinoma tissues, normal tissues and miR-7 mimics transfected HepG2 cells was detected by Western blot. The correlation of the levels of miR-7 and Raf1 mRNA was determined by Pearson correlation analysis.@*Results@#The relative expression level of miR-7 in HCC was 0.49±0.02, significantly lower than in ANT (1.21±0.05, P<0.01). The level of miR-7 was significantly correlated the tumor volume, metastasis and prognosis of HCC patients (P<0.05). The relative expression level of miR-7 in miR-7 mimics transfected HepG2 group was 12.67±0.40, significantly higher than that in blank group (P<0.01). Compared with the blank group, the A value and invasion ability of miR-7 mimics transfected group were significantly down-regulated at 48 hours and 72 hours after transfection (P<0.01). Compared with miR-7 NC group, the luciferase activity of wild-type Raf1 reporter gene in miR-7 mimics transfected group was significantly reduced (P<0.01). The relative expression of Raf1 protein in HCC was 3.15±0.10, significant higher than in ANT (0.53±0.03, P<0.01). The relative expression of Raf1 protein in miR-7 mimics transfected group was 0.24±0.01, significantly lower than in miR-7 NC group (0.98±0.02, P<0.01). Furthermore, an negative correlation was observed between the levels of miR-7 and Raf1 in HCC tissues (P<0.05).@*Conclusions@#The expression of miR-7 in HCC is significantly decreased and inversely correlated with poor survival of HCC patients. Overexpression of miR-7 can inhibit the proliferation and invasion ability of hepatocellular carcinoma cells HepG2 by downregulating Raf1 in vitro.
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Objective To investigate the prognostic value of platelet to lymphocyte ratio (PLR) for early virological response in Entecavir (ETV)-treated chronic hepatitis B (CHB) patients with genotype C infection.Methods Ninety-one genotype C CHB patients with HBV DNA≥1×105 copies/mL were treated with ETV (0.5 mg/d) for 10-13 days.The correlation between PLR and viral load decline was evaluated by Pearson or Spearman's rank correlation coefficient.Stepwise linear regression analysis was used to establish the prediction model of virological response.Receiver operating characteristic (ROC) curves were used to evaluate the predictive value of PLR for early virological response in ETV-treated patients with genotype C hepatitis B virus (HBV) infection.Results After 10-13 days of ETV treatment, HBV DNA decreased ≥1×lg copies/mL from baseline in 89 cases of the 91 patients, while HBV DNA declined ≥2×lg copies/mL in 65 patients and 4 patients achieved HBV DNA<500 copies/mL.HBV DNA decline was positively correlated with baseline PLR levels (r=0.235 09, P<0.05).After adjustment for age, gender, Hepatitis B e Antigen (HBeAg), and treatment days, HBV DNA decline was still positively correlated with baseline PLR levels (r=0.220 26, P<0.05).Area under curve (AUC) of prediction model including age , baseline aspartate transaminase (AST) and HBV DNA was 0.759 (95% CI : 0.660-0.859, P<0.01).After adding PLR to the prediction model, the AUC was 0.780 (95% CI: 0.685-0.875, P<0.01).Conclusions PLR is predictive to early virological response in ETV-treated CHB patients with genotype C infection.Higher baseline PLR level indicates a better virological response.PLR monitoring should be recommended in CHB patients with antiviral treatment in clinical practice.
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Objective To investigate the correlation between serum level of insulin growth factor 1 (IGF-1) and the degree of liver fibrosis in patients with chronic hepatitis C (CHC) complicated to type 2 diabetes mellitus (T2DM). Methods The cases were divided into CHC with T2DM (39 cases), CHC (96 cases), T2DM (60 cases), and healthy control (60 cases) groups. Their human data were collected and the fasting blood glucose, insulin and insulin-like growth factor-1 levels were detected, and the insulin resistance index ( HOMA-IR) wwer calculated. The serum levels of alanine transaminase ( ALT) , aspartate transaminase ( AST) , hepatitis C virus ( HCV) RNA load and HCV genotypes were detected simultaneously in patients with hepatitis C, and liver stiffness were measured ( LSM) by transient elastography ( TE) and aspartate aminotransferase-to-platelet ratio index ( APRI) score was performed. Results ⑴ There was no significant difference between CHC with T2DM group and CHC group in diabetes family history(P>0. 05), but two groups were significantly lower than that of T2DM group (P<0. 05). ⑵The levels of fasting insu-lin (FI) and HOMA-IR in CHC with T2DM group and T2DM group were significantly higher than those in the other two groups (P<0. 05), while the levels of IGF-1 in two groups were significantly lower than those in the CHC group, and were more lower than the control group (P<0. 05). ⑶Compared the serum ALT, AST and HCV RNA load between CHC with T2DM group and CHC group, there had no significant differ-ence (P>0. 05);however, the proportion of 1b genotype, LSM and APRI score of CHC with T2DM group were significantly higher than those in CHC group(P<0. 05). ⑷ The level of serum IGF-1 was negatively correlated with HOMA-IR, LSM and APRI in CHC with T2DM group (r= -0. 71, -0. 75, -0. 69, P<0. 01). Conclusions The degree of hepatic fibrosis and the damage of IGF-1 synthesis in CHC patients with T2DM were significantly higher than those in non T2DM patients, which might be related to the insulin resistance caused by 1b genotype HCV infection.
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Objective To investigate the effect of long hairpin RNA ( lhRNA) expression vector targeting HBV X gene ( HBx) on replication of hepatitis B virus ( HBV) and gene expression.Methods Four kinds of small interference RNAs ( siRNAs) were synthesized and lhRNA expression vectors targeting HBx were constructed.Four siRNA oligonucleotides and two lhRNA expression vectors were transfected into HepG2.2.15 cells.HBsAg, HBV DNA in culture supernatants and HBx mRNA in HepG2.2.15 cells were detected by time-resolved immunofluorometric assay, real-time quantitative PCR, and reverse transcription PCR, respectively.Negative sequence group or empty vector group was taken as the control.Independent-samples t test was performed to evaluate the inhibition effect on replication of HBV and gene expression. Results Compared with the negative control, HBsAg, HBV DNA level in culture supernatants and HBx mRNA in HepG2.2.15 cells were significantly decreased after siRNA-1 and siRNA-4 transfected at high concentrations (60 nmol/L or 90 nmol/L) (P<0.05), especially the HBsAg and HBV DNA levels in the siRNA-1 transfection group, which were significantly decreased at 24, 48 and 72 h after transfection ( P<0.05 or P <0.01 ) . Two lhRNA expression vectors ( pMD-HBxlh1 and pMD-HBxlh4 ) were successfully constructed and transfected into HepG2.2.15 cells, HBsAg and HBV DNA level in transfected cells was significantly lower than those in negative control (P<0.05).Conclusion The novel siRNA-1 is confirmed to target HBx gene and lhRNA expression vector targeting HBx can effectively inhibit the replication of HBV and expression of HBV gene.
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Objective To evaluate prognostic factors of patients with cerebral postoperative glioma treated with three-dimensional conformal radiotherapy (3DCRT). Methods62 patients with cerebral glioma who had residual tumor received 3DCRT. Factors such as gender, age, location of the glioma, pathological type, pathological grade, degree of surgical resection, combined chemotherapy, radiation dose, Karnofsky score before treatment were analyzed to show wether the impact on prognosis, Drawing survival curve with the Kaplan-Meier method, Log-rank test for univariate analysis and multivariate analysis were performed by using COX regression model.ResultsLast follow-up time was April 1, 2009. Median follow-up was 22 months, 43patients died. The 1-, 2- and 3-year overall survival rate were 65.7 %, 34.2 % and 20.5 %. In univariate analysis, the survival was related to age (P=0.008), pathological grade (P=0.016), degree of surgical resection (P=0.000), Karnofsky score before treatment (P =0.018). In COX multivariate modal, age (P =0.031), degree of surgical resection (P =0.000), Kamofsky score before treatment (P =0.035) were independent prognostic factors. ConclusionAge ＜40 years, Karnofsky score before treatment ≥70 and mostly resection are independent factors for predicting better survival of glioma patients.
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Background and purpose: Cerebral gliomas is one of the common brain tumors, and has a poor prognosis. Therefore, multidisciplinary treatment strategy has been much investigated recently. This study investigated the efficacy of 3 dimensional conformal radiotherapy (3D-CRT) concurrent with Temozolomide chemotherapy in the treatment of postoperative cerebral gliomas. Methods: Sixty-two patients with cerebral glioma who had residual tumor surgery after surgery were randomized into 3D-CRT group (radiotherapy group, n=31) and 3D-CRT concurrent chemotherapy group (chemoradiotherapy group, n=31) prospectively. All patients received a dose of 50-60Gy/25-30F/5-6 weeks by 6MV-X ray, 1fx/day, 5 times a week. Chemotherapy regimen was Temozolomide: 75 mg/(m~2·d),concomitantly with radiotherapy,followed by 150-200mg/(m~2·d)for 5 days,28 days per cycle for total 3-6 cycles. Results: The total response rate was 35.3% (11/31) in the radiotherapy group, and 61.3% (19/31) in the chemoradiotherapy group. The difference was significant (P=0.042). But no significant difference was observed in terms of survival in the two groups (P=0.263). Stratified analysis showed that patients with grade Ⅲ gliomas in chemoradiotherapy group had better prognosis than those in the radiotherapy group (P=0.043). Conclusion. 3D-CRT concurrent with chemotherapy can improve the survival of pathological grade Ⅲ gliomas.