CAJ | 학술논문

目的：探讨解整合素样金属蛋白酶8（a disintegrin and metalloprotease domain 8，ADAM8）基因对结肠癌HCT8细胞增殖及增殖信号转导途径PI3 K-Akt-mTOR的影响。方法体外培养结肠癌HCT8细胞，构建含ADAM8基因过表达质粒载体和ADAM8基因RNA干扰质粒载体；转染至HCT8细胞，分为过表达组、干扰组和对照组。采用EdU和MTS方法检测细胞增殖情况，PI3K酶活检测试剂盒检测细胞内PI3K酶活，Western Blot方法检测细胞内Akt、p-Akt相对表达量和mTOR的表达量。结果过表达组细胞增殖能力（1.22±0.13）显著高于对照组（1.00±0.12）（P＜0.05），干扰组细胞增殖能力（0.78±0.11）显著低于对照组（1.00±0.12）（P＜0.05）。与对照组相比，过表达组和干扰组细胞内 PI3 K的酶活变化差异无统计学意义。ADAM8过表达后细胞内 p-Akt 和mTOR表达量均增加，干扰表达后p-Akt和mTOR表达量均降低。结论 ADAM8可通过增强 Akt的磷酸化和 mTOR的表达促进HCT8细胞增殖。
목적：탐토해정합소양금속단백매8（a disintegrin and metalloprotease domain 8，ADAM8）기인대결장암HCT8세포증식급증식신호전도도경PI3 K-Akt-mTOR적영향。방법체외배양결장암HCT8세포，구건함ADAM8기인과표체질립재체화ADAM8기인RNA간우질립재체；전염지HCT8세포，분위과표체조、간우조화대조조。채용EdU화MTS방법검측세포증식정황，PI3K매활검측시제합검측세포내PI3K매활，Western Blot방법검측세포내Akt、p-Akt상대표체량화mTOR적표체량。결과과표체조세포증식능력（1.22±0.13）현저고우대조조（1.00±0.12）（P＜0.05），간우조세포증식능력（0.78±0.11）현저저우대조조（1.00±0.12）（P＜0.05）。여대조조상비，과표체조화간우조세포내 PI3 K적매활변화차이무통계학의의。ADAM8과표체후세포내 p-Akt 화mTOR표체량균증가，간우표체후p-Akt화mTOR표체량균강저。결론 ADAM8가통과증강 Akt적린산화화 mTOR적표체촉진HCT8세포증식。
Objective To investigate the effect of a disintegrin and metalloprotease domain(ADAM8)gene on colon cancer HCT8 cell proliferation and proliferation signal transduction pathways PI3K-Akt-mTOR. Methods Colon cancer HCT8 cells were cultured in vitro,and transfected with ADAM8 overexpression plasmid and RNA interfering plasmid. Cell proliferation were detected by EdU and MTS method assays. PI3K activity was measured by PI3K activity detection kit,Western Blot method was performed to detect the ratio of Akt and p-Akt and expression of mTOR. Results Compared with control group(1.00 ±0.12),the cell proliferation in ADAM8 overexpression group(1.22 ±0.13)was significantly higher (P<0.05)and in RNA interfering group(0.78 ±0.11)was significantly lower while PI3K activity had no significant changes in three groups. After ADAM8 overexpression,and the ratio of p-Akt/Akt and mTOR expression were increased significantly,while reduced significantly after RNA interferered. Conclusion ADAM8 can promote HCT8 cell proliferation through enhancing the phosphorylation of Akt and promoting the expression of mTOR.