摘要
目的探讨放疗调控食管癌细胞程序性死亡蛋白配体1(PD-L1)表达的可能机制。方法以不同剂量X线照射食管癌细胞株Eca109、Kyse150和TE1,同时设6 Gy照射+AG490组。采用实时荧光定量聚合酶链反应检测细胞中PD-L1 mRNA的表达,采用Western blot检测细胞中PD-L1、信号转导和转录激活因子3(STAT3)和磷酸化细胞信号传导和转录活化因子3(p-STAT3)的蛋白表达,采用酶联免疫吸附试验检测细胞中白细胞介素6(IL-6)的蛋白表达。结果食管癌细胞Eca109、Kyse150和TE1中,PD-L1 mRNA的表达量分别为2.86±0.30、960.01±21.27和106.78±6.67,均高于正常食管细胞株HET-1A(1.07±0.15,均P<0.01);PD-L1蛋白的表达量分别为0.091±0.036、1.533±0.079和0.914±0.035,亦均高于正常食管细胞株HET-1A(0.063±0.01,均P<0.01)。经6 Gy照射后,Eca109、Kyse150和TE1细胞中PD-L1蛋白的表达量峰值分别为0.135±0.007、1.66±0.06和1.32±0.06,均高于0 h组(分别为0.09±0.01、1.21±0.05和0.93±0.03,均P<0.01);p-STAT3蛋白的表达量峰值分别为1.44±0.26、0.75±0.04和1.92±0.17,均高于0 h组(分别为0.18±0.05、0.48±0.02和0.36±0.06,均P<0.01)。不同剂量照射后,Eca109、Kyse150和TE1细胞中IL-6蛋白表达均显著增加(均P<0.01)。IL-6/STAT3信号通路被特异性抑制剂AG490阻断后,6 Gy放射+AG490组Eca109、Kyse150和TE1细胞中PD-L1蛋白的表达水平分别为0.11±0.03、1.07±0.08和0.96±0.11,与相应细胞株的0 Gy照射组(分别为0.09±0.01、0.96±0.05和0.85±0.09)比较,差异均无统计学意义(均P>0.05);p-STAT3蛋白的表达水平分别为0.76±0.11、0.59±0.06和0.96±0.12,与相应细胞株的0 Gy照射组(分别为0.67±0.08、0.54±0.06和0.84±0.11)比较,差异亦均无统计学意义(均P>0.05)。结论放疗可能通过IL-6/STAT3信号通路调控食管癌细胞的PD-L1表达。
Objective To explore the possible mechanism of radiotherapy regulating the expression of PD-L1 in esophageal carcinoma.Methods Three esophageal cancer cell lines(Eca109,Kyse150,TE1)were irradiated with different doses of X-rays,and 6 Gy+AG490 group was set.The mRNA expression of PD-L1 was detected by real-time quantitative polymerase chain reaction(RT-qPCR).The protein expressions of PD-L1,STAT3,p-STAT3 were detected by western blotting and the protein level of IL-6 was detected by ELISA.Results The mRNA expressions of PD-L1 in Eca109,Kyse150 and TE1 were 2.86±0.30,960.01±21.27 and 106.78±6.67,higher than 1.07±0.15 in normal esophageal cell line HET-1A(P<0.01).The protein expressions of PD-L1 in Eca109,Kyse150 and TE1 were 0.091±0.036,1.533±0.079 and 0.914±0.035,higher than 0.063±0.01 in normal esophageal cell line HET-1A(P<0.01).After 48 hours of 6 Gy irradiation,the protein expression levels of PD-L1 in Eca109,Kyse150 and TE1 were 0.135±0.007,1.66±0.06 and 1.32±0.06,higher than 0.09±0.01,1.21±0.05 and 0.93±0.03 of the 0 Gy group(P<0.01),while the protein expression levels of p-STAT3 in Eca109,Kyse150 and TE1 were 1.44±0.26,0.75±0.04 and 1.92±0.17,higher than 0.18±0.05,0.48±0.02 and 0.36±0.06 of the 0 Gy group(P<0.01).IL-6 protein expression increased significantly after different doses of irradiation(P<0.01).After the IL-6/STAT3 signaling pathway was blocked by the specific inhibitor AG490,the expressions of PD-L1 of Eca109,Kyse150 and TE1 in the 6 Gy+AG490 groups were 0.11±0.03,1.07±0.08 and 0.96±0.11,without significant differences of 0.09±0.01,0.96±0.05 and 0.85±0.09 of the 0 Gy group(P>0.05),while the protein expressions of p-STAT3 were 0.76±0.11,0.59±0.06 and 0.96±0.12,without significant differences of 0.67±0.08,0.54±0.06 and 0.84±0.11 of the 0 Gy group(P>0.05).Conclusion Radiotherapy may regulate the expression of PD-L1 in esophageal cancer cells through IL-6/STAT3 signaling pathway.
作者
周益臣
庞学舟
朱红蕾
何宇
沈洋
马代远
Zhou Yichen;Pang Xuezhou;Zhu Honglei;He Yu;Shen Yang;Ma Daiyuan(Department of Oncology,Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,China)
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2022年第5期389-394,共6页
Chinese Journal of Oncology
基金
四川省卫生健康委员会重点课题(17ZD006)。
