ECHS1 Involved in Antagonising Apoptosis in HepG2 Cells via the Mitochondrial Pathway

目的:观察烯脂酰辅酶A水合酶短链1(EnOyl-COA HydrATASE SHOrT CHAIn 1,ECHS1)对HEPg2细胞凋亡的影响。方法:构建ECHS1基因的干扰质粒,转染HEPg2细胞后通过嘌呤霉素筛选稳定干扰ECHS1的细胞株(HEPg2-SIECHS1),利用WESTErn blOT验证其干扰效率;利用流式细胞仪及TunEl方法检测ECHS1干扰后HEPg2细胞凋亡情况,再采用WESTErn blOT检测凋亡相关蛋白表达水平。结果:成功构建了ECHS1的干扰质粒;WESTErn blOT验证了ECHS1干扰后HEPg2细胞株中ECHS1表达低于空白对照组(未转染HEPg2细胞)及阴性对照组(空载体Pu6转染HEPg2细胞)(P<0.05);流式细胞仪术显示HEPg2-SIECHS1组细胞株凋亡率(14.98±1.07)%,阴性对照组(10.55±0.19)%,两者差异有统计学意义(P<0.05);TunEl实验显示HEPg2-SIECHS1组细胞株凋亡率(6.13±0.12)%,阴性对照组(2.89±0.21)%,两者差异有统计学意义(P<0.05);与阴性对照组比较,ECHS1干扰后P53、促凋亡蛋白bAX及bId均表达增加。结论:ECHS1通过线粒体途径拮抗HEPg2细胞凋亡。Objective: To investigate the impact of enoyl-CoA hydratase short chain 1(ECHS1) on apoptosis of HepG2 cells.Methods: The ECHS1 interference plasmids were constructed and transfected into HepG2 cells.The stable interference cell lines were screened via puromycin.Interference efficacy was detected by Western blot.Flow cytometry and TUNEL assays studied the apoptosis of HepG2 cells.Apoptosis-related proteins were detected by Western blot.Results: The HepG2 cells with stable ECHS1 gene interference were successfully established.The expression level of ECHS1 protein in HepG2 cells after transfection with ECHS1 siRNA was significantly lower than that in the blank control cells(HepG2 cells without transfection) and the negative control cells(HepG2 cells transfected with pU6 vector)(P< 0.05).Apoptosis ratio of the ECHS1 siRNA group was significant higher than that of negative control group by both Flow cytometry and TUNEL assays(P<0.05).Expressions of p53,Bax and Bid in ECHS1 siRNA group were higher than those in negative control group.Conclusion: ECHS1 may antagonise the apoptosis of HepG2 cells via the mitochondrial pathway.南京军区医学科技创新课题(10MA070); 厦门市科技计划医疗卫生创新项目(3502Z20124029)资助项

Identification of differential gene expressions in colorectal cancer and polyp by cDNA microarray

Medical Technology Innovation Project of Nanjing Military [09MA066]AIM: To screen the differential expressed genes in colorectal cancer and polyp tissue samples. METHODS: Tissue specimens containing 16 cases of colorectal adenocarcinoma and colorectal polyp vs normal mucosae were collected and subjected to cDNA microarray and bioinformatical analyses. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to confirm some of the cDNA microarray data. RESULTS: The experimental data showed that eight genes were differentially expressed, most of which were upregulated in adenomatous polyp lesions. Forty-six genes expressions were altered in colorectal cancers, of which 29 were upregulated and 17 downregulated, as compared to the normal mucosae. In addition, 18 genes were similarly altered in both adenomatous polyps and colorectal cancer. qRT-PCR analyses confirmed the cDNA microarray data for four of those 18 genes: MTA1, PDCD4, TSC1 and PDGFRA. CONCLUSION: These differentially expressed genes likely represent biomarkers for early detection of colorectal cancer and may be potential therapeutic targets after confirmed by further studies. (C) 2012 Baishideng. All rights reserved

Interaction of benzo[a]pyrene with other risk factors in hepatocellular carcinoma: a case-control study in Xiamen, China

Xiamen Municipal Science and Technology Program [3502Z20073015]Purpose: Large epidemiologic studies about the relationship between benzo[a]pyrene (B[a]P) and hepatocellular carcinoma (HCC) have been limited. B[a]P diol epoxide (BPDE) is a highly reactive metabolite of B[a]P that binds covalently to form DNA adducts. We evaluated the interaction between B[a]P exposure with other risk factors in HCC, in a case-control study of 345 HCC and 961 healthy controls. Methods: Concentration of BPDE-DNA adducts in blood was determined by enzyme-linked immunosorbent assay. The interaction between BPDE-DNA adducts and other risk factors on HCC were evaluated by multivariate logistic regression analysis. Results: Mean concentration of BPDE-DNA adducts in blood of cases was significantly higher than that of the controls. The risk of HCC increased with elevated concentration of BPDE-DNA adducts (x(2) = 203.57, P-trend < .001) and the odds ratio was 7.44 (95% confidence interval, 5.29-10.45) for the first versus fourth quartile of adduct levels. The relative excess risk due to interaction between BPDE-DNA adducts and hepatitis B virus surface antigen and drinking was 34.71 and 54.92, and the attributable proportion due to interaction was 41.53% and 75.59%, respectively. Conclusions: The high level of BPDE-DNA adducts in blood is associated with HCC and that environmental exposure to B[a]P may increase the risk of HCC, especially among drinkers and populations with hepatitis B virus infection. (C) 2014 Elsevier Inc. All rights reserved