本文選題：肺腺癌 + PC細胞�。� 參考：《中華腫瘤防治雜志》2017年09期

【摘要】：目的培美曲塞是治療非鱗癌非小細胞肺癌(non-small cell lung cancer,NSCLC)的常用藥物,其對表皮生長因子受體(pithelial growth factor receptor,EGFR)敏感突變的肺腺癌是否具有放射增敏作用尚未明確。本課題研究培美曲塞在體外對EGFR 19外顯子突變的人肺腺癌細胞株P(guān)C9的放射增敏作用,并初步探討其作用機制。方法以PC9細胞作為研究對象,采用MTT法檢測培美曲塞的20%抑制濃度(20%inhibition concentration,IC_(20)),將PC9細胞分為對照組、單獨照射組、培美曲塞單藥組和培美曲塞+照射組4組;采用流式細胞術(shù)檢測培美曲塞聯(lián)合或不聯(lián)合照射對PC9細胞的凋亡率及細胞周期的影響;克隆形成實驗檢測培美曲塞對PC9細胞的放射效應(yīng),計算存活分數(shù),擬合存活曲線;蛋白質(zhì)印跡法檢測CDC20蛋白在各組的表達。結(jié)果 MTT結(jié)果顯示,PC9細胞的增殖抑制與培美曲塞呈時間-劑量依賴性,取48h的IC_(20)(0.084μmol/L)的培美曲塞作為實驗濃度。流式細胞術(shù)結(jié)果顯示,培美曲塞單藥組和單獨照射組均可增加凋亡率,分別為(7.17±1.14)%和(10.78±1.52)%,而培美曲塞+照射組具有協(xié)同增效作用,凋亡率達(29.23±1.33)%,F=227.57,P0.001;細胞周期結(jié)果顯示,對照組G_2/M期比例為(0.79±0.63)%,培美曲塞單藥組為(0.79±0.47)%,單獨照射組為(18.21±0.72)%,培美曲塞+照射組為(25.09±1.04)%,提示培美曲塞使細胞阻滯在S期,與放射線聯(lián)合作用后,S期細胞比例減少,G_2/M期的比例明顯增多,差異有統(tǒng)計學(xué)意義,F=276.85,P0.001�？寺⌒纬蓪嶒炋崾�,培美曲塞具有較好的放射增敏作用,其放射增敏比(sensitization enhancement ratio,SER)為1.41。培美曲塞聯(lián)合照射能增加細胞周期相關(guān)蛋白CDC20的表達,F=282.12,P0.001。結(jié)論培美曲塞可提高EGFR 19外顯子突變的PC9肺腺癌細胞的放射敏感性,其機制可能與照射引起CDC20的增加致對放射敏感的G_2/M期阻滯,而照射耐受的S期比例減少有關(guān)。
[Abstract]:Objective pemetrexil is a common drug in the treatment of non-small cell lung cancer (NSCLC). It is not clear whether pemetrexed has radiosensitization effect on epidermal growth factor receptor (EGFR) sensitive mutation in lung adenocarcinoma. The aim of this study was to investigate the radiosensitizing effect of pemetrexil on human lung adenocarcinoma cell line PC9 with exon 19 mutation of EGFR in vitro, and to explore its mechanism. Methods PC9 cells were used to detect 20% inhibition concentration of pemetrexide by MTT. PC9 cells were divided into 4 groups: control group, single irradiation group, pemetrexed single drug group and pemetrexed irradiation group. Flow cytometry was used to detect the apoptosis rate and cell cycle of PC9 cells induced by combined or uncombined irradiation of pemetrexide, and clone formation assay was used to detect the radiative effect of pemetrexide on PC9 cells, to calculate the survival fraction and to fit the survival curve. The expression of CDC20 protein in each group was detected by Western blot. Results the results of MTT showed that the inhibition of proliferation of PC9 cells was time-dose dependent on pemetrexide, and the concentration of pemetrexil was 48 h IC_(20)(0.084 渭 mol / L as the experimental concentration. The results of flow cytometry showed that the apoptotic rate was increased in both pemetrexide group and irradiation group, which were 7.17 鹵1.14% and 10.78 鹵1.52%, respectively, while that in pemetrexed group was synergistic, and the apoptotic rate was 29.23 鹵1.33% (P 0.001). In the control group, the ratio of G2 / M phase was 0.79 鹵0.63, that of pemetrexed group was 0.79 鹵0.47g, that of single irradiation group was 18.21 鹵0.72g, and that of pemetrexed irradiation group was 25.09 鹵1.040.The results showed that pemetrexil made the cells block in S phase, and the proportion of cells in S phase decreased in G2M phase after combined irradiation. The difference was statistically significant (P 0.001). Clone formation experiment showed that pemetrexil had better radiosensitization, and its radiosensitization ratio was 1.41%. The combined irradiation of pemetrexide could increase the expression of cell cycle associated protein CDC20. Conclusion pemetrexil can increase the radiosensitivity of PC9 lung adenocarcinoma cells with mutation of exon 19 of EGFR, and its mechanism may be related to the increase of CDC20 induced by irradiation and the arrest of G 2 / M phase of radiation sensitive to radiation, but the decrease of S phase ratio of radiation tolerance.
【作者單位】： 廣西壯族自治區(qū)人民醫(yī)院化療一區(qū);廣西壯族自治區(qū)人民醫(yī)院科研實驗中心;
【基金】：廣西科技廳項目(桂科攻0632007-1L)
【分類號】：R730.55;R734.2

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