中华消化外科杂志
中華消化外科雜誌
중화소화외과잡지
CHINESE JOURNAL OF DIGESTIVE SURGERY
2010年
4期
262-264
,共3页
刘松%张朝阳%牛卫博%王加勇%彭程%王健%牛军
劉鬆%張朝暘%牛衛博%王加勇%彭程%王健%牛軍
류송%장조양%우위박%왕가용%팽정%왕건%우군
胃肿瘤%整合素%细胞外调节蛋白激酶%5-氟尿嘧啶
胃腫瘤%整閤素%細胞外調節蛋白激酶%5-氟尿嘧啶
위종류%정합소%세포외조절단백격매%5-불뇨밀정
Gastric neoplsms%Integrins%Extracellular signal-regulated protein kinases%5-fluorouracil
目的 探讨整合素αvβ6通过细胞外调节蛋白激酶介导胃癌细胞对5-氟尿嘧啶的耐受作用.方法 胃癌AGS细胞培养至对数生长期后分4组处理.(1)对照组:向细胞悬液加入含5-氟尿嘧啶的培养基培养24 h.(2)10D5组:向细胞悬液内加入0.1 g/L鼠抗人αvβ6单克隆抗体10D5,培养6 h后换液加入含5-氟尿嘧啶的培养基培养24 h.(3)IgG2a组:加入10D5对照剂IgG2a后用5-氟尿嘧啶处理,余同10D5组.(4)PD98059组:向细胞悬液内加入含5-氟尿嘧啶和20 μmol/L细胞外调节蛋白激酶抑制剂PD98059的培养基培养24 h.MTT法检测细胞增殖,流式细胞仪检测细胞凋亡,Western blot检测Bcl-2和caspase-3蛋白的表达情况.采用单因素方差分析和LSD检验.结果 对照组、10D5组、IgG2a组和PD98059组的抑制率分别为28.1%±2.7%、84.5%±1.6%、31.4%±5.2%和86.7%±5.2%,组间比较差异有统计学意义(F=342.5,P<0.05);凋亡率分别为6.6%±1.4%、30.6%±2.4%、8.1%±1.3%和36.0%±4.0%,组间比较差异有统计学意义(F=105.4,P<0.05).caspase-3和Bcl-2的表达水平组间比较,差异有统计学意义(F=292.1,181.6,P<0.05).结论 整合素αvβ6可通过细胞外调节蛋白激酶介导胃癌细胞对5-氟尿嘧啶产生耐受.
目的 探討整閤素αvβ6通過細胞外調節蛋白激酶介導胃癌細胞對5-氟尿嘧啶的耐受作用.方法 胃癌AGS細胞培養至對數生長期後分4組處理.(1)對照組:嚮細胞懸液加入含5-氟尿嘧啶的培養基培養24 h.(2)10D5組:嚮細胞懸液內加入0.1 g/L鼠抗人αvβ6單剋隆抗體10D5,培養6 h後換液加入含5-氟尿嘧啶的培養基培養24 h.(3)IgG2a組:加入10D5對照劑IgG2a後用5-氟尿嘧啶處理,餘同10D5組.(4)PD98059組:嚮細胞懸液內加入含5-氟尿嘧啶和20 μmol/L細胞外調節蛋白激酶抑製劑PD98059的培養基培養24 h.MTT法檢測細胞增殖,流式細胞儀檢測細胞凋亡,Western blot檢測Bcl-2和caspase-3蛋白的錶達情況.採用單因素方差分析和LSD檢驗.結果 對照組、10D5組、IgG2a組和PD98059組的抑製率分彆為28.1%±2.7%、84.5%±1.6%、31.4%±5.2%和86.7%±5.2%,組間比較差異有統計學意義(F=342.5,P<0.05);凋亡率分彆為6.6%±1.4%、30.6%±2.4%、8.1%±1.3%和36.0%±4.0%,組間比較差異有統計學意義(F=105.4,P<0.05).caspase-3和Bcl-2的錶達水平組間比較,差異有統計學意義(F=292.1,181.6,P<0.05).結論 整閤素αvβ6可通過細胞外調節蛋白激酶介導胃癌細胞對5-氟尿嘧啶產生耐受.
목적 탐토정합소αvβ6통과세포외조절단백격매개도위암세포대5-불뇨밀정적내수작용.방법 위암AGS세포배양지대수생장기후분4조처리.(1)대조조:향세포현액가입함5-불뇨밀정적배양기배양24 h.(2)10D5조:향세포현액내가입0.1 g/L서항인αvβ6단극륭항체10D5,배양6 h후환액가입함5-불뇨밀정적배양기배양24 h.(3)IgG2a조:가입10D5대조제IgG2a후용5-불뇨밀정처리,여동10D5조.(4)PD98059조:향세포현액내가입함5-불뇨밀정화20 μmol/L세포외조절단백격매억제제PD98059적배양기배양24 h.MTT법검측세포증식,류식세포의검측세포조망,Western blot검측Bcl-2화caspase-3단백적표체정황.채용단인소방차분석화LSD검험.결과 대조조、10D5조、IgG2a조화PD98059조적억제솔분별위28.1%±2.7%、84.5%±1.6%、31.4%±5.2%화86.7%±5.2%,조간비교차이유통계학의의(F=342.5,P<0.05);조망솔분별위6.6%±1.4%、30.6%±2.4%、8.1%±1.3%화36.0%±4.0%,조간비교차이유통계학의의(F=105.4,P<0.05).caspase-3화Bcl-2적표체수평조간비교,차이유통계학의의(F=292.1,181.6,P<0.05).결론 정합소αvβ6가통과세포외조절단백격매개도위암세포대5-불뇨밀정산생내수.
Objective To explore the role of integrin αvβ6 in mediating the tolerance of gastric cancer AGS cells to 5-fluorouracil, and to determine whether direct β6-extracellular signal-regulated protein kinase(ERK) binding is involved. Methods Gastric cancer AGS cells in the logarithmic phase were incubated with either 5-fluorouracil for 24 hours ( control group), with 0.1 g/L of mouse anti-αvβ6 monoclonal antibody 10D5 for 6 hours and then with 5-fluorouracil for24 hours ( 10D5 group), with IgG2a and 5-fluorouracil ( IgG2a group), or with 5-fluorouracil and 20 μnol/L of ERK inhibitor PD98059 for 24 hours ( PD98059 group). Cell proliferation,apoptosis and the expression of Bcl-2 and caspase-3 protein were detected by MTT assay, flow cytometry and western blotting, respectively. All data were analysed by one-way analysis of variance and LSD test. Results The cell inhibition rates of the control group, 10D5 group, IgG2a group and PD98059 group were 28.1% ±2.7%,84.5% ± 1.6%, 31.4% ±5.2%, 86.7% ±5.2%, respectively, with a significant difference ( F = 342. 5, P ＜0.05). The apoptosis rates of the control group, 10D5 group, IgG2a group, and PD98059 group were 6.6% ±1.4%, 30.6% ± 2.4%, 8.1% ± 1.3%, 36.0% ±4.0%, respectively, with a significant difference among the four groups (F = 105.4, P ＜0.05 ). There was a significant difference in the expression of caspase-3 and Bcl-2 among the four groups (F=292.1, 181.6, P＜0.05). Conclusion Integrin αvβ6 can mediate the tolerance of gastric cancer AGS cells to 5-fluorouracil through direct β6-ERK binding.
