整合素连接激酶对幽门螺杆菌所致细胞凋亡的影响
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摘要
目的探讨整合素连接激酶对幽门螺杆菌(Hp)致胃癌细胞MGC-803凋亡的影响。方法实验分为空白对照组(胃癌细胞MGC-803正常条件培养),激动剂作用组(MGC803细胞与0.4μmol/L的整合素连接激酶激动剂LPTP共培养),Hp作用组(MGC803细胞与Hp(MOI值为100∶1)共培养)和激动剂加Hp作用组(MGC803细胞与0.4μmol/L的LPTP共培养1h,与Hp(MOI值为100∶1)共培养)。24h后流式细胞术检测细胞凋亡率,Western blot检测细胞凋亡相关蛋白PARP、Casepase-3的表达。结果激动剂作用组、Hp作用组、激动剂加Hp作用组细胞凋亡率分别为(23.45±1.484 92)%、(30.7±3.111 27)%、(16.15±1.202 08)%,与空白对照组(10.1±0.707 11)%比较,差异均有统计学意义(t值分别为-11.478,-9.131,-6.135,P<0.05);激动剂加Hp作用组与Hp作用组比较差异有统计学意义(t=6.169,P<0.05)。激动剂作用组、Hp作用组、激动剂加Hp作用组PARP蛋白相对表达量分别为0.89±0.57、1.245±0.21、0.87±0.057,与空白对照组0.6±0.06比较差异均有统计学意义(t值分别为-4.734,-13.492,-4.401,P<0.05);Casepase-3蛋白表达分别为0.78±0.35、1.32±0.04、0.87±0.45;与空白对照组0.47±0.07比较差异有统计学意义(t值分别为-5.635,-14.577,-8.037,P<0.05)。结论整合素连接激酶在Hp感染MGC-803细胞中起抗凋亡的作用。
Objective To examine the effect of integrin-linked kinase on apoptosis of the MGC-803 gastric cancer cell line induced by H.pylori. Methods MGC-803 gastric cancer cells were divided into a blank control group(cultured under normal conditions),cells cultured in the presence of LPTP(MGC803 cells cultured in the presence of 0.4μmol/L LPTP,an integrin-linked kinase agonist),cells co-cultured with H.pylori(MGC803 cells cultured with H.pylori(MOI=100:1)),cells co-cultured with H.pylori in the presence of LPTP(MGC803 cells cultured with H.pylori for1 hin the presence of 0.4μmol/L LPTP(MOI= 100:1)co-culture).After 24 h,the rate of apoptosis was detected using flow cytometry,and the level of PARP and Casepase-3 expression was detected using Western blotting. ResultsThe rate of apoptosis was(23.45.±1.48492)%in cells cultured in the presence of LPTP,(30.7±3.11127)%in cells co-cultured with H.pylori,(16.15±1.20208)%in cells co-cultured with H.pylori in the presence of LPTP.The rate of apoptosis differed significantly from that(10.1±0.70711%)in the blank control group(t=-11.478,-9.131,and-6.135;P<0.05).The rate of apoptosis differed significantly in cells co-cultured with H.pylori in the presence of LPTP and cells co-cultured with H.pylori(t=6.169,P<0.05).The level of PARP protein expression was 0.89±0.57 in cells cultured in the presence of LPTP,1.245±0.21 in cells co-cultured with H.pylori,and 0.87±0.057 in cells co-cultured with H.pylori in the presence of LPTP.The level of PARP protein expression differed significantly from that(0.6±0.06)in the blank control group(t=-4.734,-13.492,and-4.401;P<0.05).The level of Casepase-3 protein expression was 0.78±0.35 in cells cultured in the presence of LPTP,1.32±0.04 in cells co-cultured with H.pylori,and 0.87±0.45 in cells co-cultured with H.pylori in the presence of LPTP.The level of Casepase-3 protein expression differed significantly from that(0.47±0.07)in the blank control group(t=-5.663,-14.577,and-8.037;P<0.05). Conclusion Integrin-linked kinase plays an anti-aging role in MGC-803 cells infected with H.pylori.
Objective To examine the effect of integrin-linked kinase on apoptosis of the MGC-803 gastric cancer cell line induced by H.pylori. Methods MGC-803 gastric cancer cells were divided into a blank control group(cultured under normal conditions),cells cultured in the presence of LPTP(MGC803 cells cultured in the presence of 0.4μmol/L LPTP,an integrin-linked kinase agonist),cells co-cultured with H.pylori(MGC803 cells cultured with H.pylori(MOI=100:1)),cells co-cultured with H.pylori in the presence of LPTP(MGC803 cells cultured with H.pylori for1 hin the presence of 0.4μmol/L LPTP(MOI= 100:1)co-culture).After 24 h,the rate of apoptosis was detected using flow cytometry,and the level of PARP and Casepase-3 expression was detected using Western blotting. ResultsThe rate of apoptosis was(23.45.±1.48492)%in cells cultured in the presence of LPTP,(30.7±3.11127)%in cells co-cultured with H.pylori,(16.15±1.20208)%in cells co-cultured with H.pylori in the presence of LPTP.The rate of apoptosis differed significantly from that(10.1±0.70711%)in the blank control group(t=-11.478,-9.131,and-6.135;P<0.05).The rate of apoptosis differed significantly in cells co-cultured with H.pylori in the presence of LPTP and cells co-cultured with H.pylori(t=6.169,P<0.05).The level of PARP protein expression was 0.89±0.57 in cells cultured in the presence of LPTP,1.245±0.21 in cells co-cultured with H.pylori,and 0.87±0.057 in cells co-cultured with H.pylori in the presence of LPTP.The level of PARP protein expression differed significantly from that(0.6±0.06)in the blank control group(t=-4.734,-13.492,and-4.401;P<0.05).The level of Casepase-3 protein expression was 0.78±0.35 in cells cultured in the presence of LPTP,1.32±0.04 in cells co-cultured with H.pylori,and 0.87±0.45 in cells co-cultured with H.pylori in the presence of LPTP.The level of Casepase-3 protein expression differed significantly from that(0.47±0.07)in the blank control group(t=-5.663,-14.577,and-8.037;P<0.05). Conclusion Integrin-linked kinase plays an anti-aging role in MGC-803 cells infected with H.pylori.
引文
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[2]李宁,李亚斐,吴雄飞.整合素连接激酶生物学功能研究进展[J].生命的化学,2016,36(3):385-9.
[3]Zhao G,Guo LL,Xu JY,et al.Integrin-linked kinase in gastric cancer cell attachment,in vasion and tumor growth[J].World J Gastroenterol,2011,17(30):3487-96.
[4]Peroukides S,Bravou V,Varakis J,et al.ILK over-expression in human hepatocellular carcinoma and liver cirrhosis correlates with activa-tion of Akt[J].Oncol Rep,2008,20(6):1337-44.
[5]张凤梅,李胜水,李秀清,等.ILK和MMP-9在胃癌及其侵袭、转移中的表达及意义[J].胃肠病学和肝病学杂志,2015(1):95-8.
[6]李娇娇,荣倩玉,季晓飞,等.幽门螺杆菌hp0788基因对GES-1细胞功能影响的初步研究[J].中国病原生物学杂志,2017,12(7):614-8.
[7]荣倩玉,李娇娇,季晓飞,等.幽门螺杆菌在两种液体培养基中生长规律的探讨[J].中国病原生物学杂志,2017,12(8):738-40.
[8]Zhang ZW,Patchett SE,Farthing MJ.Role of Helicobacter pylori and p53in regulation of gastric epithelial cell cycle phase progression[J].Dig Dis Sci,2002(47):987-95.
[9]Gang S,Gaoliang OY,Shideng B,et al.The activation of Akt/PKB signaling pathway and cell survival[J].J Cell Mol Med,2005,9(1):59-71.
[10]Subham B,Nicholas F,Meredith S,et al.Akt phosphorylates the Yes-associated protein,YAP,to induce interaction with 14-3-3and attenuation of p73-mediated apoptosis[J].Molecular Cell,2003,11(1):11-23.
[11]Pugazhenthi S,Nesterova A,Sable C,et al.Akt/Protein kinase B up-regulates Bcl-2expression through cAMP-response elementbinding protein[J].J Biol Chem,2000,275(15):10761-6.
[12]Chen Y,Wang Y,Xu W,et al.Analyss on the mechanism of Helicobacter pylori-induced apoptosis in gastric cancer cell line BGC-823[J].Int J Mol Med,2005,16(4):741-5.
[13]Pierzchalski P,Pytko-Polonczyk J,Jaworek J,et al.Only live Helicobacter pylori is capable of casepase-3dependent apoptosis induction in gastric mucosa epithelial cells[J].J Physiol Pharmacol,2009,60(4):119.
[14]Franke TF,Yang SI,Chan TO,et al.The protein kinase encoded by the Akt proto-oncogene is a target of the PDGF-activated phosphatidylinositol 3-kinase[J].Cell,1995,81(5):727-36.
[15]Alessi DR,Andjelkovic M,Caudwell B,et al.Mechanism of activation of protein kinase B by insulin and IGF-1[J].EMBO J,1996,15(23):6541-51.
[16]Reed JC.Proapoptotic multidomain Bcl-2/Bax-family proteins:mechanisms,physiologyical roles,and therapeutic opportunities[J].Cell Death Differ,2006,13(8):1378-86.