CAJ | 학술논문

目的：探讨T790M突变所致吉非替尼耐药肺腺癌细胞株PC-9/GR放射敏感性的变化情况及经射线照射后PC-9/GR吉非替尼耐药的逆转情况。方法选取吉非替尼敏感肺腺癌细胞株PC-9及T790M突变所致吉非替尼继发性耐药肺腺癌细胞株PC-9/GR。应用液相芯片法检测这两种细胞中人第10号染色体缺失的磷酸酶及张力蛋白同源的基因（ PTEN ） mRNA 表达水平；应用免疫印迹法检测这两种细胞中磷酸化哺乳动物雷帕霉素靶蛋白（ p-mTOR）、磷酸化蛋白激酶B（ p-Akt）、PTEN蛋白表达水平；采用集落形成实验观察射线照射细胞的存活情况；应用四甲基偶氮唑盐（ MTT）比色法检测细胞的生长抑制情况。结果（1） PC-9/GR的PTEN mRNA及其蛋白表达水平均高于PC-9（P<0.05），而p-mTOR、p-Akt蛋白表达水平则低于PC-9（P<0.05）。（2）在不同剂量6 MV X线作用下，PC-9/GR的细胞存活分数（SF）均低于PC-9（P<0.05）。（3）在不同浓度吉非替尼作用下，射线照射的PC-9/GR的生长抑制率均高于未照射的 PC -9/GR （ P <0.05），半数抑制浓度（ IC50）则低于未照射的PC-9/GR（P<0.05）。射线照射的PC-9/GR每一吉非替尼浓度的生长抑制率均高于未经照射的 PC-9/GR（P<0.05）。结论在体外实验条件下，肺腺癌细胞株 PC -9/GR 的放射敏感性较其亲代 PC -9升高，射线照射后的PC-9/GR在一定程度上能逆转吉非替尼的耐药。
목적：탐토T790M돌변소치길비체니내약폐선암세포주PC-9/GR방사민감성적변화정황급경사선조사후PC-9/GR길비체니내약적역전정황。방법선취길비체니민감폐선암세포주PC-9급T790M돌변소치길비체니계발성내약폐선암세포주PC-9/GR。응용액상심편법검측저량충세포중인제10호염색체결실적린산매급장력단백동원적기인（ PTEN ） mRNA 표체수평；응용면역인적법검측저량충세포중린산화포유동물뢰파매소파단백（ p-mTOR）、린산화단백격매B（ p-Akt）、PTEN단백표체수평；채용집락형성실험관찰사선조사세포적존활정황；응용사갑기우담서염（ MTT）비색법검측세포적생장억제정황。결과（1） PC-9/GR적PTEN mRNA급기단백표체수평균고우PC-9（P<0.05），이p-mTOR、p-Akt단백표체수평칙저우PC-9（P<0.05）。（2）재불동제량6 MV X선작용하，PC-9/GR적세포존활분수（SF）균저우PC-9（P<0.05）。（3）재불동농도길비체니작용하，사선조사적PC-9/GR적생장억제솔균고우미조사적 PC -9/GR （ P <0.05），반수억제농도（ IC50）칙저우미조사적PC-9/GR（P<0.05）。사선조사적PC-9/GR매일길비체니농도적생장억제솔균고우미경조사적 PC-9/GR（P<0.05）。결론재체외실험조건하，폐선암세포주 PC -9/GR 적방사민감성교기친대 PC -9승고，사선조사후적PC-9/GR재일정정도상능역전길비체니적내약。
Objective To investigate the change of radiosensitivity in gefitinib -resistant lung adenocarcinoma cell line PC-9/GR caused by T790M mutation,and the reversal of drug resistance in gefitinib-resistant PC-9/GR after X ray irra-diation. Methods Gefitinib-sensitive lung adenocarcinoma cell line PC-9 and T790M mutation-induced secondary gefitinib-resistant lung adenocarcinoma cell line PC-9/GR cell line were selected. Liquid chip method was applied to detect the expres-sion level of PTEN mRNA in the two kinds of cells;Western blotting method was used to detect the expression level of p-mTOR,p-Akt and PTEN proteins;Colony formation assay was carried out to observe the survival situation of the cells which were irradiated by X ray;MTT colorimetric method was used to study the growth inhibition of cells. Results The expression lev-els of PTEN mRNA and PTEN proteins in PC-9/GR were significantly higher than those in PC-9(P<0. 05),while the ex-pression levels of p-mTOR and p-Akt in PC-9/GR were significantly lower than those in PC-9 ( P<0. 05 ) . With the ac-tion of different doses of 6 MV X ray,the survival fraction(SF)of PC-9/GR was significantly lower than that of PC-9(P<0. 05). With the action of different concentrations of gefitinib,the growth inhibition rate of PC-9/GR with irradiation was sig-nificantly higher than that of unirradiated PC-9/GR(P<0. 05),and the IC50 values in irradiated PC-9/GR were significantly lower than those in non-irradiated PC-9/GR(P<0. 05). In every concentration of gefitinib,the growth inhibition rates of PC-9/GR with irradiation were significantly higher than those of unirradiated PC-9/GR(P<0. 05). Conclusion Under the experimental conditions in vitro,the radiosensitivity of lung adenocarcinoma PC-9/GR cells is higher than that of its parental PC-9 cells,and the irradiated PC-9/GR cells can reverse gefitinib resistence to some extent.