1.Expression of stomach cancer-associated transcript 16 in gastric cancer tissues and its effects on gastric cancer biological behavior
Jianfeng ZHANG ; Qianfeng ZHANG ; Xiaoling KUAI ; Zhongshang SUN ; Zhenbiao MAO
Chinese Journal of Digestion 2017;37(5):313-318
Objective To study the expression of long non-coding RNA (LncRNA) stomach cancer-associated transcript 16 (STCAT16) in gastric cancer tissues and its effects on the proliferation, migration and invasion of gastric cancer cells.Methods The different expression of STCAT16 in 32 cases of gastric carcinoma and corresponding adjacent tissues was detected by real-time fluorescence quantitative polymerase chain reaction (PCR).The STCAT16 overexpression plasmid and empty vector was separately transfected gastric cancer cell line AGS with low expression of STCAT16.The cell proliferation of empty vector group, non-transfection group and STCAT16 analogue transfection group was evaluated by cell counting kit-8 (CCK-8) assay at 0, 24, 48, 72 and 96 hours after transfection.The colony forming ability was tested by colony formation assay.The cell invasion ability was measured by Transwell chamber assay and migration ability was tested by scratch-wound assay.The effects of STCAT16 on tumorigenicity in vivo were verified by tumorigenicity experiments in nude mice.T-test and one-way analysis of variance were performed for data analysis.Repeated measures analysis of variance was used to compare the repeated measured data among groups.Chi square test and Fisher exact probability method were used for comparison of counting data.Results The expression of STCAT16 in gastric cancer tissues was low (0.87±0.19), while it was high in corresponding adjacent tissues (2.32±0.37), and the difference was statistically significant (t=-20.859, P<0.05).The expression of STCAT16 of STCAT16 analogue transfection group was higher than that of empty vector group (3.43±0.25 vs 1.00±0.06), and the difference was statistically significant (t=-16.795,P<0.05).Compared to empty vector group and non-transfection group, the cell proliferation decreased in STCAT16 analogue transfection group at 72 and 96 hours after transfection (1.41±0.07, 1.42±0.08, 1.03±0.09, and 1.72±0.11, 1.78±0.14, 1.24±0.08, respectively), and the differences were statistically significant (t=15.043,5.358, 12.193 and 8.109, all P<0.05).The results of colony formation assay indicated that the colony forming ability of gastric cells in STCAT16 analogue transfection group was lower than that in empty vector group (97.3±9.1 vs 185.0±20.1) and non-transfection group (97.3±9.1 vs 138.0±11.1), and the differences were statistically significant (t=11.634 and 4.417,both P<0.05).The results of Transwell assay showed that the number of AGS cells passing through the membrane of STCAT16 analogue transfection group was significantly less than those of empty vector group and non-transfection group (151.0±28.1 vs 228.0±38.2 and 151.0±28.1 vs 199.3±17.9), and the differences were statistically significant (t=4.823 and 4.747,both P<0.05).After transfection for 48 hours and 72 hours, the scratch-wound repair rate of STCAT16 analogue transfection group decreased, compared with those of empty vector group and non-transfection group ((52.67±6.11)%, (53.33±5.51)%, (42.67±4.72)%, and (90.67±2.51)%, (90.60±5.41)%, (69.67±1.52)%, respectively), and the differences were statistically significant (t=5.773, 5.955, 21.000 and 5.881, all P<0.05).The results of tumorigenicity in nude mice showed that compared with those of empty vector group, the tumor size of STCAT16 analogue transfection group was smaller at one-, two-, three-and four-week ((0.42±0.10) cm3 vs (0.16±0.05) cm3, (0.66±0.13) cm3 vs (0.34±0.05) cm3, (1.25±0.22) cm3 vs (0.54±0.13) cm3, (2.54±0.46) cm3 vs (0.78±0.41) cm3)), and the differences were statistically significant (t=3.175, 4.190, 7.996 and 9.705, all P<0.05).Conclusions STCAT16 is lowly expressed in gastric cancer tissues.The proliferation, migration, invasion ability and tumorigenicity in nude mice of gastric cancer cell is inhibited after upregulating the expression of STCAT16.
2.Expression of galectin 3 and its effects on proliferation and invasion in pancreatic cancer SW1990 cells
Guoxiong ZHOU ; Miao ZHANG ; Xiaoling DING ; Haifeng ZHANG ; Wei CAO ; Lishuai JU ; Xiaoling KUAI ; Hong ZHANG ; Zhengfu XU
Chinese Journal of Pancreatology 2012;12(5):320-322
Objective To investigate the expression of galectin-3 in pancreatic cancer cell and its effect on the proliferation and invasion of SW1990.Methods Immunocytochemistry and semi-quantitative RT-PCR was used to detect the expression of galectin-3 protein and mRNA in SW1990,PANC1 and ASPC-1 cell lines.Galectin-3 mono-antibody of different concentrations ( 1,2,3,5 μg/ml) was used to treat SW1990 cells for 24,48,72 h,CCK-8 kits were used to detect the proliferation in SW1990 cells; Transwell chamber was used to study the invasion in SW1990.Results Expression of galectin-3 protein and mRNA was present in SW1990,PANC1,and ASPC-1.Galectin 3 mono-antibody inhibited the proliferation and number of invasive cells in a dose and time dependant manner.The inhibitory rates at 72 h were 19.8%,29.9% and 42.7% in 2,3,5 μg/ml galectin 3 mono-antibody groups,the difference among them and control group was statistically significant ( P < 0.05 ).The inhibite rate of permeating membrane cells in 3 μg/ml galectin-3 mono antibody was 37.1%,the difference between this group and control group was statistically significant (P <0.05 ).ConcLusions Galectin-3 is highly expressed in pancreatic cancer cells.Galectin-3 antibody can inhibit the proliferation and migration capability of SW1990 cells.
3.Effect of microRNA-32 on the proliferation and migration of gastric cancer cells
Xiaoling KUAI ; Mengjiao SONG ; Zhihua YU ; Jianfeng ZHANG ; Xiaoqi CHEN ; Hong ZHANG ; Zhenbiao MAO
Chinese Journal of Digestion 2013;33(7):465-469
Objective To explore the effect of microRNA-32 (miRNA-32)on the biological behaviors of gastric cancer cell and its mechanism.Methods Gastric cancer cell line SGC-7901 cells were transiently transfected with miRNA-32 analogue,miRNA-32 inhibitor and empty plasmid vectors by lipofectamine and divided into analogue transfection group,inhibitor transfection group,empty plasmid transfection group and non-transfection group.The expression of green fluorescent protein was observed under fluorescent microscopy.The expression of miRNA-32 at mRNA level was detected by quantificational real-time polymerase chain reaction.The cell proliferation was evaluated by CCK-8 assay.The cell migration ability was measured by scratch test and Transwell chamber assays.The data were analyzed by one-way ANOVA.Results Compared with empty plasmid transfection group and non-transfection group,the expression of miRNA-32 mRNA of miRNA-32 analogue transfection group (relative quantitative value:2.327) was significantly up-regulated and that of miRNA-32 inhibitor transfection group (relative quantitative value:0.402) was significantly down regulated (F=11.238,P<0.05).The width of scratch of miRNA-32 analogue transfection group was (61.39± 2.21) μm at 24 hours; miRNA-32 inhibitor transfection group was (29.97±0.66) μm.The migration distance of inhibitor transfection group was far than that of analogue transfection group (F=9.371,P<0.05).After transfection for 48 hours,the cell number of migrated cells of analogue transfection group was significantly less than that of non transfection group,which was 16.93±4.63 and 93.93± 7.09,respectively (F=6.853,P<0.05).After transfection for 48 hours and 72 hours,the cell growth inhibiting rate of miRNA 32 analogue transfection group was (43.474 ± 18.636)% and (45.050±23.764)%,respectively,the cell growth was significantly inhibited (F=7.986 and 8.635,P=0.028 and 0.032).Conclusion The cell growth and migration ability of human gastric cancer cell line SGC-7901 are obviously inhibited through upregulating the expression of miRNA-32.
4.Analysis of Risk Factors of Recurrent Acute Pancreatitis
Xiaoling KUAI ; Haifang DING ; Jianfeng ZHANG ; Zhaoxiu LIU ; Chengqi GUAN ; Zhenbiao MAO
Chinese Journal of Gastroenterology 2016;21(12):729-732
Background:Recurrent acute pancreatitis(RAP)is a special type of acute pancreatitis(AP). Finding the cause is the key to avoid the recurrence of RAP. Aims:To investigate the risk factors of RAP. Methods:The clinical data of 43 patients with RAP and 130 patients with only one time AP(control group)were retrospectively analyzed. Risk factors of recurrence of RAP were analyzed. Results:Univariate analysis showed that no significant differences in etiology,severity, serum levels of cholesterol,calcium,white blood cell count,amylase,ALT,AST,total bilirubin,direct bilirubin and C-reactive protein were found between RAP group and control group(P > 0. 05). Serum triglyceride level,blood glucose level and CT score in RAP group were significantly higher than those in control group(t = 3. 260,P < 0. 05;t = 2. 720, P < 0. 05;t = 2. 162,P < 0. 05). Logistic regression analysis demonstrated that serum triglyceride level,blood glucose level and CT score were risk factors of RAP(oR = 1. 86,95% CI:1. 05-3. 68,P = 0. 03;oR = 1. 23,95% CI:1. 01-1. 50,P = 0. 04;oR = 2. 46,95% CI:1. 00-6. 03,P = 0. 04). Conclusions:The recurrence of RAP is related with serum triglyceride level,blood glucose level and CT score.
5.Correlation between the expression of sodium borate transporter member 11 of solute carrier family 4 protein and tumor protein P53 protein and clinicopathological features and prognosis in gastric carcinoma
Yue QI ; Shuang LIU ; Mengjing SUN ; Peng LI ; Huijun ZHU ; Xiaoling KUAI
Chinese Journal of Digestion 2018;38(12):811-817
Objective To explore the expression and clinical significance of sodium borate transporter member 11 of solute carrier family 4 (SLC4A11) and tumor protein P53 (TP53) proteins in gastric cancer (GC).Methods From March 2004 to December 2009,a total of 415 patients with GC,who received operation at Department of General Surgery of Affiliated Hospital of Nantong University,were enrolled.The clinical data and gastric tissues samples of them were collected.At same period,64 patients with non-malignant gastric diseases,who underwent surgery at Department of General Surgery of Affiliated Hospital of Nantong University,were also recruited.The clinical data and gastric precancerous tissues were also collected.The tissues were maken into tissue microarray (TMA).The expression of SLC4A11 and TP53 protein in tissue microarrays was detected by immunohistochemical staining.The relationship between the two proteins and clinicopathological parameters and prognosis of patients were analyzed.Chi square test,and univariate and multivariate analyses were used for statistical analysis.Results The positive rates of protein expression of SLC4A11 and TP53 in GC tissues were 48.19% (200/415) and 34.94% (145/415),respectively,which were higher than 17.19% (11/64) and 6.25% (4/64) in gastric precancerous lesions,respectively,and the differences were statistically significant (x2 =24.150 and 59.345;both P<0.01).The results of statistical analysis showed that the expression of SLC4A11 was correlated with human epidermal growth factor receptor 2(Her2) levels,depth of invasion,distant metastasis and TNM staging (x2 =28.056,11.300,8.880,24.943;all P<0.05).Meanwhile,the expression of TP53 was related with depth of invasion,lymph node metastasis,distant metastasis,TNM staging and preoperative carcinoembryonic antigen (CEA;x2=12.333,7.875,9.347,20.307,10.678;all P<0.05).The expression of TP53 was significantly positive correlated with SLC4A11 expression (x2 =6.237,P=0.013).The results of univariate analysis showed that SLC4A11,TP53,SLC4A11+/TP53+,Her2,preoperative CEA and cancer antigen 19-9 (CA19-9) levels were correlated with the poor prognosis of GC patients (hazard ratio (HR)=1.947,1.459,1.797,1.419,2.221,1.908;all P<0.05),while the results of multivariate analysis indicated that positive SLC4A11 and TNM staging were the independent parameters for judging the prognosis of patients with GC (HR =1.954,1.468,both P<0.05).Conclusions The high expression of SLC4A11 and TP53 is related to the occurrence and development of GC.Combined detection of their changes will contribute to the early diagnosis and prognostic evaluation of patients with GC.