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Relationship between autophagic process and PARP-1 in xenograft nasopharyngeal carcinoma CNE-2 cells in nude mice following radiotherapy[Objective]To investigate whether radiation can cause autophagy,and explore the relationship between PARP-1 and radiation-induced autophagy in transplanted tumors of CNE-2 cells in nude mice.[Methods]1.The CNE-2 cells were inoculated subcutaneously in nude mice and divided into 2 groups: 0Gy group and 10 Gy group.The 10 Gy group received 10 Gy irradiation dose.The PARP-1,LC3 II and P62 in tumor xenografts of nude mice were detected by western blot,to determine whether radiation can cause autophagy in xenograft tumors in nude mice.2.PARP-1 gene silenced CNE-2 was inoculated into nude mice and divided into CNE-2 group,NC group and PARP-1-shRNA group.The expression of PARP-1 was detected by RT-qPCR and Western blot after tumor formation,to verify the silent effect of PARP-1.3.In the CNE-2 group,NC group,and PARP-1-shRNA group,nude mice received 10 Gy irradiation dose.Western blot was used to detect the expression of PARP-1,LC3 II and P62,and the relationship between PARP-1 and autophagy was confirmed.[Results]1.Compared with 0Gy group,the expression of PARP-1 and LC3 II in 10 Gy group increased,and the expression of P62 decreased.The difference was statistically significant(P<0.05).2.Compared with CNE-2 group and NC group,the expression of PARP-1 in PARP-1-shRNA group was decreased,and the difference was statistically significant(P<0.05).3.After 10 Gy irradiation,compared with CNE-2 group and NC group,the expressions of PARP-1 and LC3 II in PARP-1-shRNA group decreased,and the expression of P62 increased,the difference was statistically significant(P<0.05).[Conclusion]Radiotherapy can cause autophagy in transplanted tumors of CNE-2 cells in nude mice,and PARP-1 is the upstream regulatory point of autophagy.Part ? PARP-1 mediates autophagy in nasopharyngeal carcinoma CNE-2 cells xenograft nude mice by AMPK/mTOR pathway following radiotherapy[Objective] To investigate whether PARP-1 regulates autophagy in nude mice transplanted tumors of CNE-2 cells through AMPK/mTOR pathway.[Methods] Nude mice were divided into 0Gy group,10 Gy group,AICAR+10Gy group,Compound C+10Gy group,and PARP-1-sh RNA+10Gy group.The latter 4 groups received 10 Gy irradiation dose.AICAR +10Gy group and Compound C +10Gy group received intraperitoneal injection of AICAR and Compound C,respectively.PARP-1-sh RNA+10Gy group inoculated with PARP-1 silenced CNE-2 cells.The expression of PARP-1,p-AMPK,AMPK,p-mTOR,mTOR,p-P70S6 K,P70S6K,LC3 II and P62 were detected by Western blot.[Results] 1.Compared with 0Gy group,the expression of PARP-1,p-AMPK/AMPK,and LC3 II in 10 Gy group increased,and the expression of p-mTOR/mTOR,p-P70S6K/P70S6 K and P62 decreased.The difference was statistically significant(P<0.05).2.Compared with 10 Gy group,the expression of p-AMPK/AMPK and LC3 II in AICAR+10Gy group increased,while the expression of p-mTOR/mTOR,p-P70S6K/P70S6 K and P62 decreased.The difference was statistically significant(P<0.05).There was no significant difference in PARP-1 expression(P>0.05).3.Compared with 0Gy group,there was no significant difference in the expression of p-AMPK/AMPK,LC3 II,p-mTOR/mTOR,p-P70S6K/P70S6K and P62 between Compound C +10Gy group and PARP-1-sh RNA + 10 Gy group(P>0.05).The expression of PARP-1 in Compound C +10Gy group increased,the difference was statistically significant(P<0.05),while the expression of PARP-1 in PARP-1-sh RNA+10Gy group had no significant difference(P>0.05).[Conclusion] After radiotherapy,PARP-1 regulates autophagy in nude mice transplanted with CNE-2 cells through the AMPK/mTOR pathway.Part ? Effect of interference with PARP-1-AMPK/mTOR pathway on radiosensitivity of nasopharyngeal carcinoma CNE-2 cells transplanted in nude mice[Objective] To investigate the effects of interfering with PARP-1-AMPK/mTOR signaling pathway on radiosensitivity of transplanted tumors in nude mice.[Methods] Nude mice were divided into 0Gy group,PARP-1-sh RNA group,Compound C group,10 Gy group,Compound C +10Gy group,PARP-1-sh RNA + 10 Gy group.The last three groups received a dose of 10 Gy.The tumor growth curve and tumor weight were measured.The expression of PARP-1 and Bax protein was detected by immunohistochemistry and western blot.[Results] 1.Compared with 0Gy group,the volume of tumor volume in 10 Gy group,PARP-1-sh RNA group and Compound C group was slower and the weight was lighter,the difference was statistically significant(P<0.05);Compared with 10 Gy group,PARP-1-sh RNA group,and Compound C group,the tumor volume growth rate of PARP-1-sh RNA+10Gy group and Compound C+10Gy group was slower and lighter,and the difference was statistically significant(P <0.05).2.Compared with 0Gy group,the expression of PARP-1 in 10 Gy group and Compound C+10Gy group increased,the difference was statistically significant(P<0.05);the expression of Bax protein in 10 Gy group,PARP-1-sh RNA group and Compound C group increased,the difference was statistically significant(P<0.05).Compared with 10 Gy group,Bax expression in PARP-1-sh RNA+ 10 Gy group and Compound C+10Gy group increased,and the difference was statistically significant(P<0.05).[Conclusion] Interfering with the PARP-1-AMPK/mTOR pathway can increase the radiosensitivity of transplanted tumors in nude mice with nasopharyngeal carcinoma CNE-2 cells. |
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