AZD5363对胆管癌细胞的作用及其机制研究

An exprimental study on AZD5363 suppressing cholangiocarcinoma cells

目的探讨Akt信号通路抑制剂AZD5363对胆管癌细胞增殖及侵袭的影响。方法将一种新的毗咯并嘧啶衍生的化合物AZD5363作用于QBC939和RBE胆管癌细胞系，应用Westernb10t检测Akt及下游蛋白表达及mTOR蛋白表达。用CCK-8实验检测药物对细胞增殖和毒性作用。采用Transwell法检测癌细胞侵袭能力。结果AZD5363作用于QBC9393细胞半数致死量药物浓度（24±9）与空白对照组比较，差异有统计学意义（f=4．47，P〈0．05），RBE细胞半数致死量药物浓度（21±8）与空白对照比较差异有统计学意义（t=4．41，P〈0．05）。QBC939肿瘤的侵袭能力在药物浓度为20μmol／L和μmol／L迁移细胞数分别是（63±12）和（271±27），差异有统计学意义。胆管癌细胞RBE在药物20μmol／L（58±23）和0μmol／L（235±21）的细胞计数，差异有统计学意义。AZD5363抑制Akt及其下游分子的磷酸化。促进QBC939细胞的mTOR分子的磷酸化。结论AZD5363抑制胆管癌细胞的增殖和侵袭能力，可能与抑制Akt及其下游分子的磷酸化，促进QBC939细胞roTOR的磷酸化有关。

Objective To investigate the effect of Akt pathway inhibitor AZD5363 on cell proliferation and invasion of QBC939 and RBE cholangiocarcinoma cells and the mechanism. Methods Western blotting was used to detect Akt and downstream protein and roTOR protein expression in two cancer cell lines after process by AZD5363. Inhibition rate and cytotoxieity was tested by CCK-8 assay, and Transwell assay was used to evaluate the invasive ability of cancer cells. Results QBC9393 cell exposed to AZD5363 LD50 drug concentration （ 24± 9 ） was significantly different compared with control group （t = 4.47, P 〈 0. 05 ） , RBE cells LD50 drug concentration （ 21 ± 8 ） was significantly different compared with control （t =4. 41, P 〈 0. 05）. Tumor invasion capacity of QBC939 in drug concentrations of 20 μmoL/L （ 63 ± 12 ） and 0 μmol/L （ 271± 27 ）, the difference was statistically significant. RBE exposed AZD5363 upon drug concentrations of 20 μmol/L （58 ±23 ） and 0 μmol/L （235 ± 21 ）, the difference was statistically significant. AZD5363 promotes phosphorylation of mTOR in QBC939. Conclusions AZD5363 inhibits the proliferation and migration, inhibiting the phosphorylation of Akt and its downstream molecules. AZD5363 promotes phosphorylation of roTOR in QBC939.