中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2012年
8期
577-581
,共5页
蔡嵘%任刚%丁夏%王以政%金冶宁
蔡嶸%任剛%丁夏%王以政%金冶寧
채영%임강%정하%왕이정%금야저
胃肿瘤%瞬时受体电势通道6%细胞增殖%细胞周期%小鼠
胃腫瘤%瞬時受體電勢通道6%細胞增殖%細胞週期%小鼠
위종류%순시수체전세통도6%세포증식%세포주기%소서
Stomach neoplasms%TRPC6%Cell proliferation%Cell cycle%Mice
目的 探讨瞬时受体电势通道6(TRPC6)基因对胃癌细胞增殖的作用及其分子机制.方法 采用Western blot法和免疫组化方法检测30例胃癌及癌旁组织中TRPC6蛋白的表达水平.以腺病毒载体介导的TRPC6显性负突变体抑制胃癌AGS细胞内源性TRPC6通道后绘制细胞生长曲线,检测单细胞克隆形成能力和裸鼠移植瘤的皮下成瘤能力；流式细胞仪检测细胞周期；Western blot 法检测Cdc2磷酸化水平.结果 TRPC6蛋白在胃癌及癌旁组织中的相对表达量分别为(21.60±8.32)％和(7.14±2.24)％,差异有统计学意义(P＜0.05).转染24、48、72、96h后,与Ad-GFP转染组相比,Ad-DNC6转染组的细胞增殖率分别下降(36.90+1.13)％、(44.06±2.17)％、(52.12±2.76)％和(50.89±1.97)％.Ad-DNC6转染组和Ad-GFP转染组胃癌AGS细胞的克降形成率分别为(14.70+3.00)％和(43.80±7.00)％.在转染0、24、36、48 h后,Ad-DNC6转染组和Ad-GFP转染组的G2/M期细胞比例分别为(20.34±1.98)％、(24.31±2.37)％、(27.70±2.36)％、(35.10±3.07)％和(18.40+2.01)％、(18.30±1.72)％、(17.50±1.74)％、(16.80±1.71)％.抑制TRPC6通道能明显抑制裸鼠移植瘤的生长(P＜0.05).结论 TRPC6通道通过凋节细胞周期G2期的进程来影响胃癌细胞的增殖.
目的 探討瞬時受體電勢通道6(TRPC6)基因對胃癌細胞增殖的作用及其分子機製.方法 採用Western blot法和免疫組化方法檢測30例胃癌及癌徬組織中TRPC6蛋白的錶達水平.以腺病毒載體介導的TRPC6顯性負突變體抑製胃癌AGS細胞內源性TRPC6通道後繪製細胞生長麯線,檢測單細胞剋隆形成能力和裸鼠移植瘤的皮下成瘤能力；流式細胞儀檢測細胞週期；Western blot 法檢測Cdc2燐痠化水平.結果 TRPC6蛋白在胃癌及癌徬組織中的相對錶達量分彆為(21.60±8.32)％和(7.14±2.24)％,差異有統計學意義(P＜0.05).轉染24、48、72、96h後,與Ad-GFP轉染組相比,Ad-DNC6轉染組的細胞增殖率分彆下降(36.90+1.13)％、(44.06±2.17)％、(52.12±2.76)％和(50.89±1.97)％.Ad-DNC6轉染組和Ad-GFP轉染組胃癌AGS細胞的剋降形成率分彆為(14.70+3.00)％和(43.80±7.00)％.在轉染0、24、36、48 h後,Ad-DNC6轉染組和Ad-GFP轉染組的G2/M期細胞比例分彆為(20.34±1.98)％、(24.31±2.37)％、(27.70±2.36)％、(35.10±3.07)％和(18.40+2.01)％、(18.30±1.72)％、(17.50±1.74)％、(16.80±1.71)％.抑製TRPC6通道能明顯抑製裸鼠移植瘤的生長(P＜0.05).結論 TRPC6通道通過凋節細胞週期G2期的進程來影響胃癌細胞的增殖.
목적 탐토순시수체전세통도6(TRPC6)기인대위암세포증식적작용급기분자궤제.방법 채용Western blot법화면역조화방법검측30례위암급암방조직중TRPC6단백적표체수평.이선병독재체개도적TRPC6현성부돌변체억제위암AGS세포내원성TRPC6통도후회제세포생장곡선,검측단세포극륭형성능력화라서이식류적피하성류능력；류식세포의검측세포주기；Western blot 법검측Cdc2린산화수평.결과 TRPC6단백재위암급암방조직중적상대표체량분별위(21.60±8.32)％화(7.14±2.24)％,차이유통계학의의(P＜0.05).전염24、48、72、96h후,여Ad-GFP전염조상비,Ad-DNC6전염조적세포증식솔분별하강(36.90+1.13)％、(44.06±2.17)％、(52.12±2.76)％화(50.89±1.97)％.Ad-DNC6전염조화Ad-GFP전염조위암AGS세포적극강형성솔분별위(14.70+3.00)％화(43.80±7.00)％.재전염0、24、36、48 h후,Ad-DNC6전염조화Ad-GFP전염조적G2/M기세포비례분별위(20.34±1.98)％、(24.31±2.37)％、(27.70±2.36)％、(35.10±3.07)％화(18.40+2.01)％、(18.30±1.72)％、(17.50±1.74)％、(16.80±1.71)％.억제TRPC6통도능명현억제라서이식류적생장(P＜0.05).결론 TRPC6통도통과조절세포주기G2기적진정래영향위암세포적증식.
Objective To investigate the essential role and mechanism of TRPC6 gene in the development of gastric cancer.Methods The expression of TRPC6 protein was assessed in gastric cancer tissues and normal tissues adjacent to the cancer from 30 patients with gastric cancer.The inhibiting effect of TRPC6 activity on cell growth,cell cycle of a human gastric cancer cell line AGS cells,tumor progression and development of xenografted human gastric cancer in a mouse model was tested using dominant-negative mutant TRPC6 ( DNC6 ). The survival of mice bearing xenografted tumors in the GFP and DNC6 was compared using Kaplan-Meier analysis.All statistical tests were two-sided.Results The TRPC6 protein in the tumor tissues and pars-tumor tissues was ( 21.60 ± 8.32) ％ versus ( 7.14 ± 2.24 ) ％.After transfection of DNC6 virus for 24 hours,48 hours,72 hours and 96 hours,the growth inhibition rates of gastric cancer cells were (36.90±1.13)％,(44.06±2.17)％,(52.12±2.76)％ and (50.89±1.97)％,respectively.The clone formation rates of control group and DNC6 group were ( 14.70 ± 3.00)％ versus (43.80 ± 7.00)％.After transfection with DNC6 virus for 0,24,36 and 48 hours,the G2/M phase arrest was (20.34 ±1.98)％,(24.31 ±2.37)％,(27.70 ±2.36)％,(35.10 ±3.0)％ in the DNC6 group and (18.40 ±2.01 ) ％,( 18.0％ ± 1.72 ) ％,( 17.50 ± 1.74) ％,( 16.80 ± 1.71 ) ％ in the control group,respectively.Inhibition of TRPC6 activity also reduced the subcutaneous tumor volume in the mouse models with xenografted human tumors (P ＜ 0.05 ).Conclusion In the preclinical models tested,TRPC6 channels are essential for gastric cancer development via regulation of G2/M phase transition.
