Genetic susceptibility of gastric cancer with single nucleotide polymorphisms(SNPs) of apoptotic genes - caspase-3 and caspase-7 / 实用肿瘤学杂志

Objective The SNPs of caspase family have been studied in breast cancer,head and neck cancer,esophageal cancer,lung cancer and other cancer. There are few studies between the SNP of CASP3,7 and the risk of gastric cancer in Chinese population. The aim of this study was to investigate the relationship between the SNP of CASP3,7 genes and the genetic susceptibility of gastric cancer using large samples. Methods Blood samples from 1000 cases of gastric cancer and 1036 cases of normal non-cancer control in Northeast China were collected for genomic DNA extraction. Based on the dbSNP NCBI database and HapMap data-base,potential SNP sites were selected for CASP3,7,which locate CASP3,CAS′s 3′UTR. The Tasman probe method was used for gen-otyping of the SNP sites of CASP3,7. The difference between the different variables in the case-control group was analyzed by the bi-lateral χ2 test. After the Hardy-Weinberg genetic balance test for each locus,the χ2 test was used to compare the genotype frequency differences between the case and control groups. Univariate and multivariate logistic regression models were used to analyze the associ-ation between genotype and disease. The stratification analysis of each locus compared the genotypes between the different sexes,ages, smoking,and drinking status. Results The χ2 test compared the differences between the case and control groups. The results showed that there was a significant difference in the smoking,drinking status and smoking in packet number( Pack-years) between the case and control groups(P<0. 05). The genotype frequencies of selected loci were in accordance with Hardy-Weinberg′s law of genetic balance(P>0. 05). Combining and analyzing genotypes with risk alleles,there showed that individuals with two risk genotypes in- creased by 69. 6% in comparison with individuals with 0~1 risk genotype. After adjusted for covariate effects,individuals with three risk genotypes were increased by 27. 6% when compared to individuals with 0~1 risk genotypes. Individuals with more than one risk genotype were increased by 35% when compared to individuals with 0-1 risk genotypes. In the stratified analysis,after combination of two genes,the risk genotype in the sub-layer of age ≤60 years old,male,never smoking,annual smoking package ≤25,gastric non-cardiac adenocarcinoma was statistically associated with disease,i. e. more risk of gastric cancer. Conclusion Univariate analy-sis showed that the SNPs at the four sites selected in this study were not associated with the risk of gastric cancer. However,multivari-ate analysis of CASP3 and CASP7 at the four sites showed that individuals with two risk genotypes increased the risk of gastric cancer in comparison with individuals with 0~1 risk genotypes. In addition,after stratified analysis of the two sites of CASP7,the risk of gas-tric cancer is more obvious in people aged≤60 years,never smoking or smoking≤25 packs per year,and non-gastric cardia adeno-carcinoma.

Effects of Shenfu Qiangxin Pills on the Expression of LC3b and Bax in Myocardial Cells of Rats with Car-diorenal Syndrome / 中国药房

OBJECTIVE:To study the effects of Shenfu qiangxin(SFQX)pills on the expression of autophagy-associated pro-tein LC3b and pro apoptotic gene Bax in myocardial cells of rats with cardiorenal syndrome (CRS). METHODS:Rats were ran-domly divided into sham operation group(water),model group(water),positive control group(Captopril tablets 2.3 mg/kg)and SFQX pills high-dose,medium-dose and low-dose groups [13.2,6.6,3.3 g(crude drug)/kg],with 10 rats in each group. CRS mod-el was induced in those groups by abdominal-aortae-constriction+acute renal ischemia reperfusion injury except for sham operation group;and they were given relevant medicine intragastrically 8 week after operation,once a day,for consecutive 4 weeks. Plasma contents of Cr and ALD,the protein expression of LC3b and Bax in myocardial tissue of rats were detected 24 h after last medica-tion;ventricular index was calculated,and morphological change of myocardial tissue was observed. RESULTS:Compared with sham operation group,the plasma contents of Cr and ALD,ventricular index and the protein expression of LC3b in myocardial tis-sue increased significantly in model group (P<0.05 or P<0.01);and myocardial cell suffered from endochylema red deletion, myocardial cross striation disorder,intercellular space fibrosis aggravation and so on. Compared with model group,the plasma con-tents of Cr and ALD(except for positive control group)and the protein expression of LC3b and Bax in myocardial tissue decreased significantly in positive control group and SFQX pills high-dose group(P<0.05 or P<0.01);myocardial pathological change was improved;the ventricular index decreased significantly in SFQX pills low-dose and medium-dose groups (P<0.05). CONCLU-SIONS:SFQX pills can decrease the plasma contents of Cr and ALD,inhibit myocardial cell autophagy and apoptosis in CRS rats.

The altered expression of apoptosis-related genes and its clinical significance by siRNA-Ku70 silencing in drug-resistant lung cancer cell line / 中国免疫学杂志

Objective:To investigate the effect of Ku70 silencing on the expression of apoptosis-related genes in human NSLC cell line,A549 and drug-resistant A549DDP.Methods:Expression of Ku70mRNA in human A549 cells was examined by RT-PCR.Expression of apoptosis-related genes was examined by PCR-Array assay.Results: The Ku70mRNA was higher in A549DDP cells than in A549 cells.The expression of BAX,TP53 and TP73 was significantly increased whereas the expression of TNFRSF1A and BFAR decreased in siRNA-Ku70 A549DDP cells when compared to those of A549DDP cells.Conclsion:The amount of Ku70 mRNA is higher in A549DDP cells,suggesting that Ku70 is important for drug-resistance; Silencing Ku70 might reverse the drug-resistance in A549DDP cells through regulation of expression of apoptofic genes.

Role of Apoptosis in Acute Rejection of Pancreaticoduodenal Transplantation in Rats / 中国普外基础与临床杂志

Objective To investigate the roles of cell apoptosis and the gene expressions of Fas, FasL, bcl-2 and bax in acute rejection of pancreaticoduodenal transplantation and to evaluate the function of duodenum biopsy for early detection of rejection in rats. Methods Wistar and SD rats were divided into two groups: ①Wistar rats that underwent allogenic pancreaticoduodenal transplantation from the organs of SD rats; ②Wistar rats that received homogenic transplantation. The grafts were then harvested on day 3, 5 and 7 after the transplantation, and all graft samples were observed with HE staining and TUNEL was also used to detect apoptotic cells. The expressions of Fas, FasL, bcl-2 and bax were measured by immunochemical method. According to Nakhleh’s score, pathologic features of transplanted pancreas and duodenum were ranged from one to three scores in order. Results The percentage of same or different scores between the pathological scores of pancreas and duodenum in allogenic pancreaticoduodenal transplantation group were 61.1% (11/18) and 38.9% (7/18) respectively, and there were 6 specimens of pancreatic tissue got higher scores with only one higher score for duodenum. There were significant differences of histopathologic rejection scores and apoptotic indices between the two groups, respectively (P

The Effect of Prostaglandin E1 on Apotosis in Ischemic Skin Island Flap of Rats

Apoptosis is a physiologic or programmed cell death process which is controlled by genes and it is essential for the function and the appropriate development of multicellular organism. Apoptosis is also thought to be one of the main mechanisms of cell death in ischemic tissues. The effect of prostaglandin E1(PGE1) is proven to be useful in the recovery of ischemic changes by inducing vasodilation of peripheral vessels and platelet disaggregation. Prostaglandin is also known to suppress apoptosis in a serum deprived cell. The purpose of this study is to evaluate the effects of PGE1 on the apoptosis in the ischemic skin island flap. Thirty Sprague-Dawley rats were used. In control group(n=15), 3x5cm sized skin island flap based on the superficial epigastric vessel was elevated and its pedicle was occluded for 14 hours. After removing the vessel clamp, skin flap was reperfused for 5 hours and harvested. In experimental group(n=15), a ischemic skin island flap was also made as in the control group except the interarterial administration of the PGE1 right after elevation of the flap and after removing the clamp. H&E, TUNEL and immunohistochemical stains for p53 and bax proteins were performed. There were ischemic changes in gross and microscopic findings in both groups. Immunohistochemical staining for p53 protein shows many positive cells with nuclear staining in squamous epithelium of the control group, but sparse positive cells in the experimental group. Immunohistochemical stainings for bax protein shows many positive cells with cytoplasmic staining in squamous epithelium of the control group, but sparse positive cells in the experimental group. The apoptotic index was significantly lower in the experimental group(2.39+/-1.76(p=0.0001)) than in the control group(7.53+/-2.05). These data indicate that PGE1 suppresses the apoptosis in the ischemic skin island flap.

Expression of Apoptotic Genes in Mouse Preimplantation Embryo Development / 대한불임학회지

OBJECTIVES: The aim of this study was to evaluate the influence of three different media on preimplatation embryo development and the expression of Bcl-2, Mcl-1, Bax, and Bok in mouse. MATERiALS AND METHODS: Two-cell embryos were retrieved from iCR female mice (4 weeks old) at 48 hr after hCG injection and cultured in Ham's F-10, HTF, and G1.2 media. The developmental rate of 2-cell embryos was evaluated from 24 hr to 72 hr after culture. RT-PCR was performed for the detection of Bcl-2, Mcl-1, Bax, and Bok gene expression. RESULTS: The rates of morula and blastocyst in HTF and G1.2 media (88%, 98.1%) were significantly higher than those in Ham's F-10 media (39.6%) at 48 hr. Likewise, the rates of hatching and hatched blastocyst in HTF and G1.2 media (21.9%, 52.9%) were higher than those in Ham's F-10 media (3.5%) at 72 hr. Bcl-2 and Bax mRNAs were highly detected in embryos cultured in Ham's F-10 when compared in embryos cultured in HTF and G1.2. in contrast, the expression of Mcl-1 and Bok was not significantly different. CONCLUSION: These results show that HTF and G1.2 culture media increase the rate of blastocyst formation and stimulate Bcl-2 and Bax gene expression in mouse preimplantation embryos.

CHANGES IN mRNA EXPRESSIONS OF bcl-2、bax、p53 AND ICE GENE IN CEREBRAL TISSUS OF RATE EXPOSED TO REPEATED+Gz / 解放军医学杂志

The purpose of this study is to observe the changes in gene expressions of bcl 2, bax, p53 and interleukin 1? converting enzyme (ICE) in the cerebral tissue of rat exposed to repeated +Gz, and to explore the pathogenetic role of apoptosis in brain damage induced by repeated +Gz exposures. The expression levels of bcl 2, bax, p53 and ICE in cerebral tissue of rats exposed to repeated +Gz were measured by semi quantitative reverse transcription polymerase chain reaction (RT PCR), and the apoptotic cells in brain tissue were detected by terminal deoxynuleotidyl transferase mediated dUTP nick end labeling technique. The results showed that the bcl 2 expression levels in the brain 6h and 24h after repeated +Gz exposures were significantly lower than those of control group, whereas the bax, p53 and ICE expression levels in the brains tissue 6h and 24h after repeated +Gz exposures were significantly higher than those of control group. Apoptotic cells could be observed in cerebral cortex, CA1 subregion of hippocampus and striatum at 6h and 24h after repeated +Gz exposures. It is suggested that the changes in bcl 2, bax, p53 and ICE expressions in rat brain can be induced by repeated +Gz exposures and apoptosis might be one of the molecular mechanisms of brain damage induced by repeated +Gz exposures