沉默IGF1R表达对胃癌细胞侵袭与迁移能力的影响及机制
|
摘要
目的探讨胰岛素样生长因子1受体(IGF1R)表达量降低对胃癌细胞侵袭、迁移能力的影响及其作用机制。方法体外培养胃癌GC9811-P细胞,取对数生长期细胞,采用RNA干扰技术进行GC9811-P细胞转染。干扰组细胞转染靶向IGF1R的小干扰RNA(IGF1-siRNA);阴性组细胞转染siRNA-NC;对照组细胞不做任何处理。Transwell法检测细胞的迁移、侵袭能力,Western Blot检测转染细胞中IGF1R、丝氨酸/苏氨酸蛋白激酶(AKT)、磷酸化AKT(p-AKT)、磷脂酰肌醇-3激酶(PI3K)、磷酸化PI3K(p-PI3K)蛋白的相对表达水平。结果干扰组细胞中IGF1R蛋白的相对表达水平显著低于对照组和阴性组(P均<0.05)。干扰组细胞的迁移、侵袭数目显著低于对照组和阴性组(P均<0.05)。对照组、阴性组、干扰组细胞中AKT、PI3K蛋白的表达水平无显著差异(P均>0.05)。干扰组细胞中p-AKT、p-PI3K蛋白的表达水平较对照组和阴性组降低,差异有统计学意义(P均<0.05)。结论沉默IGF1R的表达可以降低胃癌细胞的侵袭、迁移能力,其作用机制可能是通过影响经典PI3K/AKT信号通路发挥作用。
Objective To investigate the effect of lowering insulin-like growth factor 1 receptor( IGF1 R) expression on invasion and migration abilities of gastric cancer cells and its mechanism. Methods The gastric cancer GC9811-P cells were cultured in vitro,and logarithmic phase cells were used for the transfection of GC9811-P cells by RNA interference technique. The cells in interfering group were transferred by small interference RNA targeting IG1 R( IGF1-siRNA). The cells in negative group were transferred by siRNA-NC. No any treatment was given for the cells in the control group.Transwell test was used to detect cell invasion and migration abilities. Western Blot method was used to detect the relative expression levels of IGF1 R,serine/threonine protein kinase( AKT),phosphorylated AKT( p-AKT),phosphoinositide3-kinase( PI3 K) andphosphorylated PI3 K( p-PI3 K) proteins in transferred cells. Results The relative expression level of IGF1 R protein in cells in interfering group was significantly lower than those in cells of negative group and control group( all P < 0. 05). The number of cells of invasion and migration in interfering group was significantly lower than those in cells of negative group and control group( all P < 0. 05). There were no significant differences in the expression levels of AKT and PI3 K proteins among three groups( all P > 0. 05). The relative expression levels of p-AKT and p-PI3 K proteins in cells of interfering group were significant lower than those in cells of negative group and control group( all P < 0. 05).Conclusion Silencing IGF1 R expression can decrease the invasion and migration abilities of gastric cancer cells,and the mechanism may be through the classic PI3 K/AKT signaling pathway.
Objective To investigate the effect of lowering insulin-like growth factor 1 receptor( IGF1 R) expression on invasion and migration abilities of gastric cancer cells and its mechanism. Methods The gastric cancer GC9811-P cells were cultured in vitro,and logarithmic phase cells were used for the transfection of GC9811-P cells by RNA interference technique. The cells in interfering group were transferred by small interference RNA targeting IG1 R( IGF1-siRNA). The cells in negative group were transferred by siRNA-NC. No any treatment was given for the cells in the control group.Transwell test was used to detect cell invasion and migration abilities. Western Blot method was used to detect the relative expression levels of IGF1 R,serine/threonine protein kinase( AKT),phosphorylated AKT( p-AKT),phosphoinositide3-kinase( PI3 K) andphosphorylated PI3 K( p-PI3 K) proteins in transferred cells. Results The relative expression level of IGF1 R protein in cells in interfering group was significantly lower than those in cells of negative group and control group( all P < 0. 05). The number of cells of invasion and migration in interfering group was significantly lower than those in cells of negative group and control group( all P < 0. 05). There were no significant differences in the expression levels of AKT and PI3 K proteins among three groups( all P > 0. 05). The relative expression levels of p-AKT and p-PI3 K proteins in cells of interfering group were significant lower than those in cells of negative group and control group( all P < 0. 05).Conclusion Silencing IGF1 R expression can decrease the invasion and migration abilities of gastric cancer cells,and the mechanism may be through the classic PI3 K/AKT signaling pathway.
引文
[1]Ferlay J,Soerjomataram I,Dikshit R,et al.Cancer incidence and mortality worldwide:sources,methods and major patterns in GLOBOCAN 2012[J].Int J Cancer,2015,136(5):E359-E386.
[2]Habib R,Akhtar J,Taqi M,et al.Lentiviral vector-mediated survivin shRNA delivery in gastric cancer cell lines significantly inhibits cell proliferation and tumor growth[J].Oncol Rep,2015,34(2):859-867.
[3]李晨阳,张文彦,张祎,等.慢病毒介导的IGF-1R基因沉默对肝细胞癌的增殖和迁移的影响[J].临床和实验医学杂志,2017,16(11):1056-1060.
[4]万璟,李小毛,舒珊荣,等.慢病毒介导的靶向沉默胰岛素样生长因子1型受体的siRNA对人子宫内膜癌细胞迁移和侵袭能力的影响[J].中国病理生理杂志,2012,28(8):1352-1357.
[5]别彩群,黄秋燕,颜英,等.转染沉默IGF1R基因的肝癌细胞株增殖、迁移及侵袭能力观察[J].山东医药,2016,56(11):1-4.
[6]崔笠,郝博,王敏,等.IGF-1R表达下调抑制肾癌786-0细胞的增殖、迁移、侵袭和细胞周期G1/S期转变[J].现代肿瘤医学,2017,25(8):1208-1213.
[7]李妍,孟凡东,付立业,等.沉默AFAP-1L2对胃癌MKN-28细胞的生物学影响[J].现代肿瘤医学,2016,24(11):1691-1694.
[8]赵明,王喆,王福光,等.IGF-1R抑制剂OSI-906对鼻咽癌细胞SUNE-1放射增敏作用的研究[J].现代肿瘤医学,2017,25(2):167-171.
[9]王悦超,亓文骞,赵平.胰岛素样生长因子1受体在胰腺癌治疗中的机制及进展[J].临床肝胆病杂志,2017,33(4):790-794.
[10]Li H,Xu L,Li C,et al.Ubiquitin ligase Cbl-b represses IGF-I-induced epithelial mesenchymal transition via ZEB2 and microRNA-200c regulation in gastric cancer cells[J].Mol Cancer,2014,13:136.
[11]张巍,吴杨,龚玲,等.靶向抑制胰岛素样生长因子Ⅰ型受体对裸鼠恶性胸腔积液的治疗作用[J].中华肿瘤杂志,2016,38(8):565-571.
[12]Haisa M.The type 1 insulin-like growth factor receptor signalling system and targeted tyrosine kinase inhibition in cancer[J].J Int Med Res,2013,41(2):253-264.
[13]Rahmani M,Aust MM,Attkisson E,et al.Dual inhibition of Bcl-2and Bcl-x L strikingly enhances PI3K inhibition-induced apoptosis in human myeloid leukemia cells through a GSK3-and bim-dependent mechanism[J].Cancer Res,2012,73(4):1340-1351.
[14]Fresno Vara JA,Casado E,De CJ,et al.PI3K/Akt signalling pathway and cancer[J].Cancer Treat Rev,2004,30(2):193-204.
[15]Cho TM,Kim WJ,Moon SK.AKT signaling is involved in fucoidaninduced inhibition of growth and migration of human bladder cancer cells[J].Food Chem Toxicol,2014,64:344-352.
[16]周炳刚,魏昌晟,张颂,等.苦参碱逆转人乳腺癌MCF-7/ADR细胞株耐药性及对PI3K/AKT信号通路下游因子的影响[J].现代肿瘤医学,2017,25(1):9-13.
[17]Zhang X,Jiang G,Sun M,et al.Cytosolic THUMPD1 promotes breast cancer cells invasion and metastasis via the AKT-GSK3-Snail pathway[J].Oncotarget,2017,8(8):13357-13366.
[18]Dai J,Qian C,Su M,et al.Gastrokine-2 suppresses epithelial mesenchymal transition through PI3K/AKT/GSK3βsignaling in gastric cancer[J].Tumour Biol,2016,37(9):12403-12410.
[19]吴爱兵,黎明春,麦宗炯,等.CK2α通过PI3K/Akt/GSK-3β信号通路调控肺腺癌A549细胞的侵袭及迁移[J].中国肺癌杂志,2017,20(4):233-238.
[20]Yan XD,Yao M,Wang L,et al.Overexpression of insulin-like growth factor-I receptor as a pertinent biomarker for hepatocytes malignant transformation[J].World J Gastroenterol,2013,19(36):6084-6092.
[2]Habib R,Akhtar J,Taqi M,et al.Lentiviral vector-mediated survivin shRNA delivery in gastric cancer cell lines significantly inhibits cell proliferation and tumor growth[J].Oncol Rep,2015,34(2):859-867.
[3]李晨阳,张文彦,张祎,等.慢病毒介导的IGF-1R基因沉默对肝细胞癌的增殖和迁移的影响[J].临床和实验医学杂志,2017,16(11):1056-1060.
[4]万璟,李小毛,舒珊荣,等.慢病毒介导的靶向沉默胰岛素样生长因子1型受体的siRNA对人子宫内膜癌细胞迁移和侵袭能力的影响[J].中国病理生理杂志,2012,28(8):1352-1357.
[5]别彩群,黄秋燕,颜英,等.转染沉默IGF1R基因的肝癌细胞株增殖、迁移及侵袭能力观察[J].山东医药,2016,56(11):1-4.
[6]崔笠,郝博,王敏,等.IGF-1R表达下调抑制肾癌786-0细胞的增殖、迁移、侵袭和细胞周期G1/S期转变[J].现代肿瘤医学,2017,25(8):1208-1213.
[7]李妍,孟凡东,付立业,等.沉默AFAP-1L2对胃癌MKN-28细胞的生物学影响[J].现代肿瘤医学,2016,24(11):1691-1694.
[8]赵明,王喆,王福光,等.IGF-1R抑制剂OSI-906对鼻咽癌细胞SUNE-1放射增敏作用的研究[J].现代肿瘤医学,2017,25(2):167-171.
[9]王悦超,亓文骞,赵平.胰岛素样生长因子1受体在胰腺癌治疗中的机制及进展[J].临床肝胆病杂志,2017,33(4):790-794.
[10]Li H,Xu L,Li C,et al.Ubiquitin ligase Cbl-b represses IGF-I-induced epithelial mesenchymal transition via ZEB2 and microRNA-200c regulation in gastric cancer cells[J].Mol Cancer,2014,13:136.
[11]张巍,吴杨,龚玲,等.靶向抑制胰岛素样生长因子Ⅰ型受体对裸鼠恶性胸腔积液的治疗作用[J].中华肿瘤杂志,2016,38(8):565-571.
[12]Haisa M.The type 1 insulin-like growth factor receptor signalling system and targeted tyrosine kinase inhibition in cancer[J].J Int Med Res,2013,41(2):253-264.
[13]Rahmani M,Aust MM,Attkisson E,et al.Dual inhibition of Bcl-2and Bcl-x L strikingly enhances PI3K inhibition-induced apoptosis in human myeloid leukemia cells through a GSK3-and bim-dependent mechanism[J].Cancer Res,2012,73(4):1340-1351.
[14]Fresno Vara JA,Casado E,De CJ,et al.PI3K/Akt signalling pathway and cancer[J].Cancer Treat Rev,2004,30(2):193-204.
[15]Cho TM,Kim WJ,Moon SK.AKT signaling is involved in fucoidaninduced inhibition of growth and migration of human bladder cancer cells[J].Food Chem Toxicol,2014,64:344-352.
[16]周炳刚,魏昌晟,张颂,等.苦参碱逆转人乳腺癌MCF-7/ADR细胞株耐药性及对PI3K/AKT信号通路下游因子的影响[J].现代肿瘤医学,2017,25(1):9-13.
[17]Zhang X,Jiang G,Sun M,et al.Cytosolic THUMPD1 promotes breast cancer cells invasion and metastasis via the AKT-GSK3-Snail pathway[J].Oncotarget,2017,8(8):13357-13366.
[18]Dai J,Qian C,Su M,et al.Gastrokine-2 suppresses epithelial mesenchymal transition through PI3K/AKT/GSK3βsignaling in gastric cancer[J].Tumour Biol,2016,37(9):12403-12410.
[19]吴爱兵,黎明春,麦宗炯,等.CK2α通过PI3K/Akt/GSK-3β信号通路调控肺腺癌A549细胞的侵袭及迁移[J].中国肺癌杂志,2017,20(4):233-238.
[20]Yan XD,Yao M,Wang L,et al.Overexpression of insulin-like growth factor-I receptor as a pertinent biomarker for hepatocytes malignant transformation[J].World J Gastroenterol,2013,19(36):6084-6092.