Objective To explore the effect and mechanism of sleeve gastrectomy （SG） for type 2 diabetes mellitus （T2DM） in Goto-Kakizaki （GK） rats. Methods Thirteen male GK rats at 12 weeks of age were randomly divided into SG group （n=7） and sham operation group （SO group， n=6）， receiving SG surgery and sham operation respectively.Body weight， food intake in 24hours， fasting plasma glucose， plasma glucagon-like peptide-1 （GLP-1）， and plasma Ghrelin of rats in 2 groups were measured or tested before operation， 1， 4， 10， and 26 weeks after operation. In 10 weeks after operation， fecal energy content of rats in 2 groups was tested， in addition， oral glucose tolerance test （OGTT） and insulin tolerance test （ITT） were performed to investigate the glucose tolerance and insulin sensitivity. Results ①Body weight：there were no significant difference on body weight between the 2 groups （P＞0.05）. Compared with time point of before operation， the body weight of both 2 groups decreased in 1 week after operation （P＜0.01）， but increased in 10 weeks and 26 weeks （P＜0.01）. ②Food intake in 24 hours：compared with SO group， the food intake of SG group were lower in 4 weeks and 10 weeks after operation （P＜0.05）. Compared with time point of before operation， the food intake of SG group were lower in 1， 4， and 10 weeks after operation （P＜0.05）， but lower only in 1 week in SO group （P＜0.05）. ③Value of fasting glucose：compared with SO group， the value of fasting glucose in SG group were lower after operation （P＜0.01）. Compared with time point of before operation， the value of fasting glucose of SG group were lower after operation （P＜0.01）， but decreased in 1 week only in SO group （P＜0.01）. ④Level of serum GLP-1：compared with SO group， the levels of serum GLP-1 in SG group were higher in 4， 10， and 26 weeks after operation （P＜0.05）. Compared with time point of before operation， the levels of serum GLP-1 in SG group were higher in 4， 10， and 26 weeks after operation （P＜0.05）， but levels of serum GLP-1 in SO group didn’t change significantly （P＞0.05）. ⑤Level of serum Ghrelin：compared with SO group， the levels of serum Ghrelin in SG group were lower at alltime points after operation （P＜0.01）. Compared with time point of before operation， the levels of serum Ghrelin in SGgroup were lower at all time points after operation （P＜0.001）， but levels of serum Ghrelin in SO group didn’t change significantly （P＞0.05）. ⑥Areas under curves （AUC）：the AUC of OGTT and ITT test in SG group were both lower than those of SO group （P＜0.01）. Conclusion SG surgery can induce the level of fasting plasma glucose， and canimprove glucose tolerance and insulin sensitivity with significant changes of levels of plasma GLP-1 and Ghrelin， sugg-esting that SG surgery may be a potential strategy to treat patient with T2DM but without obesity or insulin resistance.
Objective To identify the expression of pleiotrophin （PTN） mRNA and protein in the colorectal cancer tissues， and to explore the clinical value of it. Methods The expressions of PTN mRNA and protein in colorectal cancer tissues （colorectal cancer group） as well as normal colorectal tissues （normal control group） were tested by using in-situ hybridization and immunohistochemistry. Results The positive rates of PTN mRNA 〔63.9% （53/83） vs. 40.7%（22/54）〕 and protein〔60.2%（50/83） vs. 33.3%（18/54）〕 in colorectal cancer group were all significantly higher than those of normal control group （P=0.008， P=0.002）. There were no significant relationship between expressions of PTN mRNA and protein with gender， age， and type of tumor （P＞0.05）， but in tissues of Ⅲ-Ⅳ stage and poor differentiation，the positive rates of PTN mRNA and protein were all higher （P＜0.05）. Conclusions The over expressions of PTN mRNA and protein in colorectal cancer tissues may directly related to the invasion and metastasis. Meanwhile， it can be used as an index to predict metastasis and prognosis of colorectal cancer.
Objective To investigate effect of hepatocyte growth factor (HGF) after lentivirus-mediated RNA interference (RNAi) targeting c-Met on invasion of colonic carcinoma cell line SW480. Methods The experiment was assigned into 3 groups: NC group, the normal cells were infected by the shRNA negative control virus (the NC-20 andNC-40 represented the negative group which were added 20 ng/mL and 40 ng/mL respectively HGF after being infected); KD group, the normal cells were infected by the shRNA-c-Met target virus (the KD-20 and KD-40 represented the interfered group which were added 20 ng/mL and 40 ng/mL HGF respectively after being infected; KD1, KD2, KD3, and KD4 represented the different RNAi targets for the purpose gene); CON group, the normal cells were not infected by any virus. The lentiviral vector shRNA-c-Met was constructed and verified by polymerase chain reaction (PCR) and DNA sequencing. The SW480 cells were infected with the shRNA-c-Met after packed with lentivirus plasmid. Fourty-eight hours transfection later, the c-Met mRNA of the transfected SW480 cell was detected by real time PCR and the c-Met protein was examined by Western blot. Seventy-two hours after transfection, the cell apoptosis was detected by flow cytometry and the invasions in the different cells with stable transfection were detected by Transwell test. Results The RNAi sequence targeting c-Met gene was successfully inserted into the lentiviral vector. The shRNA-c-Met transfection resulted in an obviously reduced expression of c-Met mRNA in the SW480 cells. The efficency of gene knock down of the KD4 (the cells with No.4 target spot knocked down) was 81.4%. The shRNA-c-Met tansfection resulted in an obviously reduced expression of c-Met protein in the SW480 cells. After transfection, the apoptosis rate of the KD group was significantly higher than that in the NC group (P<0.001) or the CON group (P<0.001). The invasion ratios in the NC group, NC-20 group, and NC-40 group were significantly higher than those in the KD group (P<0.001), KD-20 group (P=0.015), and KD-40 group (P=0.017), respectively; which in the NC-20 group and NC-40 group were increased as compared with the NC group (P<0.001,P<0.001), and in the NC-40 group was increased as compared with the NC-20 group (P=0.005). The invasion ratios in the KD-20 group and KD-40 group were increased as compared with the KD group (P<0.001,P<0.001), and in the KD-40 group was increased as compared with the KD-20 group (P=0.014). Conclusion Lentivirus-mediated RNAi targeting c-Met could effectively suppress expression of c-Met in SW480 cells and could reduce invasion of HGF on SW480 cells with knocked down c-Met.
ObjectiveTo investigate the inhibitory effect of short hairpin RNA (shRNA) mediated contactin-1 (CNTN1) gene silencing on growth of human breast cancer cell line MDA-MB-468 transplanted tumors in nude mice.MethodsEighteen nude mice (4-week-old male BALB/c) were randomly equally divided into three groups: blank control group, empty vector group, and silencing group. The MDA-MB-468 cells (blank control group), MDA-MB-468 cells transfected by nonsense shRNA (empty vector group), and MDA-MB-468 cells transfected by shRNA (silencing group) were collected in the logarithmic growth period, respectively. The subcutaneous tumor models of nude mice were prepared by the subcutaneous injection of the different group cells. The tumor growth was observed and the expressions of CNTN1 and Ki-67 proteins in the transplanted tumor were detected by the immunohistochemistry.ResultsThe xenograft models of human breast cancer cells were established successfully. The tumor growth in the silencing group was significantly slower than that of the other two groups at every 3 d point (P<0.05). The tumor volume and the tumor weight in the silencing group were significantly smaller or slighter than those of the other two groups at day 18 (P<0.05). The positive rates of CNTN1 and Ki-67 protein expressions in the tumor tissues of the silencing group were lower than those of the other two groups (P<0.05), respectively.ConclusionSilencing expression of CNTN1 gene might inhibit growth of breast cancer cell line MDA-MB-468 transplanted tumors in mude mice.
ObjectiveTo detect the expression of contactin-1 (CNTN1) gene in the breast cancer tissues and explore the relation between the expression and prognosis of patients with breast cancer. MethodsThe clinicopathologic data of 35 patients with breast cancer who met the inclusion criteria from January to June in 2015 in the Shaanxi Provincial Cancer Hospital were retrospectively collected. The Western blot and immunohistochemistry methods were used to detect the CNTN1 protein expressions in the 35 breast cancer tissues and their corresponding tissues adjacent to cancer and the reverse transcriptase-PCR method was used to detect the CNTN1 mRNA expression. The relation between the CNTN1 protein expression and 5-year overall survival (OS) was analyzed. ResultsThe positive rate and expression level of CNTN1 protein in the breast cancer tissues were higher than those in the tissues adjacent to cancer [65.7% (23/35) vs. 45.7% (16/35), χ2=8.235, P=0.003; 0.825±0.221 vs. 0.412±0.117, t=12.556, P<0.001], and the expression level of CNTN1 mRNA in the breast cancer tissue was also higher than that in the corresponding tissues adjacent to cancer (0.763±0.218 vs. 0.353±0.135, t=11.162, P<0.001). The positive rates of CNTN1 protein were higher in the patients with larger tumor diameter (>2 cm), lower differentiation, later TNM stage (stage Ⅲ+Ⅳ) and axillary lymph node metastasis (P<0.05). The median OS of 35 patients with breast cancer was 47.3 months, which was 36.2 months and 52.5 months in the patients with high CNTN1 expression and low CNTN1 expression (median expression of CNTN1 protein was 0.785 as critical value) respectively (χ2=4.052, P=0.049). ConclusionPreliminary results of this study suggest that overexpression of CNTN1 gene might play an important role in occurrence and progression of primary breast cancer and is related to prognosis of survival.
Objective To detect expression of Ras association domain family 1A （RASSF1A） gene in the colonic carcinoma tissue and to analyze the relationship of this expression to its clinical features. Methods Immunohistochemistry and Western blot methods were employed for detecting the RASSF1A protein expressions in 34 colonic carcinoma tissues and corresponding normal colon tissues. RT-PCR was employed for detecting RASSF1A mRNA expression. Results ①The RASSF1A protein expression in the colonic carcinoma tissues was significantly lower than that in the normal colontissues by using immunohistochemistry〔35.3% （12/34） versus 97.1% （33/34）， P＜0.05〕.There were significant relati-onships of RASSF1A protein expressions to the tumor differentiation and TNM stage （P＜0.05）， in other words， the positive rates of RASSF1A protein in the moderately and well differentiated andⅠ＋Ⅱof TNM colonic carcinoma tissues were all higher （P＜0.05）. ② The RASSF1A protein expression in the colonic carcinoma tissues was significantly lower than that in the normal colon tissues by using Western blot 〔0.316 8±0.019 6 versus 0.914 4±0.177 6， P＜0.05〕， which was close to the result of RT-PCR〔0.158 9±0.223 7 versus 0.572 3±0.193 9， P＜0.05〕. Conclusions Absentexpre-ssion of RASSF1A gene in the colonic carcinoma tissue might play an important role in tumor genesis and tumor progre-ssion， and it might become useful early detection of the colonic carcinoma.