en Radiation Biology Radiation Biology تحقيق بديع Original Research <div style="text-align: justify;"><span style="font-size:10pt"><span style="text-justify:newspaper"><span style="text-kashida-space:50%"><span style="line-height:119%"><span style="font-family:Calibri"><span style="color:black"><span lang="en-US" style="font-size:9.0pt"><span style="font-family:Calibri"><span style="color:#1f497d"><span style="font-style:italic"><span style="font-weight:bold"><span style="language:en-US">Background</span></span></span></span></span></span><span lang="en-US" style="font-size:9.0pt"><span style="font-family:Calibri"><span style="color:#1f497d"><span style="font-weight:bold"><span style="language:en-US">:</span></span></span></span></span> <span lang="en-US" style="font-size:9.0pt"><span style="font-family:Calibri"><span style="language:en-US">To investigate the effect of DIRAS2 on the response to ionizing radiation (IR) and the related potential molecular mechanism in human ccRCC cells. </span></span></span><span lang="en-US" style="font-size:9.0pt"><span style="font-family:Calibri"><span style="color:#1f497d"><span style="font-style:italic"><span style="font-weight:bold"><span style="language:en-US">Materials and Methods:</span></span></span></span></span></span><span lang="en-US" style="font-size:9.0pt"><span style="font-family:Calibri"><span style="language:en-US"> In this paper, the expression levels of DIRAS2 in human ccRCC and paired normal tissues were obtained from the Oncomine platform and The Cancer Genome Atlas (TCGA) database, which was further validated by immunohistochemistry. DIRAS2-overexpression cell lines were constructed using a lentivirus-mediated gene expression system. A clonogenic assay was performed to evaluate cell radiation resistance. The effect of DIRAS2 on autophagy was determined by immunoblotting and immunofluorescence analysis. The expression of DIRAS2 and related signalling molecules was evaluated by immunoblotting. </span></span></span><span lang="en-US" style="font-size:9.0pt"><span style="font-family:Calibri"><span style="color:#1f497d"><span style="font-style:italic"><span style="font-weight:bold"><span style="language:en-US">Results: </span></span></span></span></span></span><span lang="en-US" style="font-size:9.0pt"><span style="font-family:Calibri"><span style="language:en-US">Here, we found that the expression of DIRAS2 was upregulated in human ccRCC. Overexpression of DIRAS2 promoted radiation resistance in ccRCC cells and enhanced the levels of radiation-induced autophagy. Moreover, inhibition of autophagy by chloroquine (CQ) pretreatment largely eliminated the effect of DIRAS2 overexpression on radiation resistance. Finally, molecular mechanism investigation showed that DIRAS2 activated the mitogen-activated protein kinase (MAPK) kinase 4 (MKK4)-c-Jun NH2-terminal kinase 1 (JNK1)-Bcl-2 pathway. </span></span></span><span lang="en-US" style="font-size:9.0pt"><span style="font-family:Calibri"><span style="color:#1f497d"><span style="font-style:italic"><span style="font-weight:bold"><span style="language:en-US">Conclusion: </span></span></span></span></span></span><span lang="en-US" style="font-size:9.0pt"><span style="font-family:Calibri"><span style="language:en-US">Taken together, these results indicated that DIRAS2 may confer radiation resistance to human RCC via autophagy induction through the MKK4-JNK1-Bcl-2 signalling pathway.</span></span></span></span></span></span></span></span></span></div> Clear cell renal cell carcinoma, DIRAS2, autophagy, radiation resistance. 805 813 http://ijrr.com/browse.php?a_code=A-10-1-1120&slc_lang=en&sid=1 C.F. Cai 7900319475328460025216 7900319475328460025216 No Department of Urology, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan 250014, Shandong, China Y. He 7900319475328460025217 7900319475328460025217 No Laboratory of Basic Medical Sciences, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan 250014, Shandong, China D. Yue 7900319475328460025218 7900319475328460025218 No Department of Urology, Affiliated Hospital of Jining Medical University, Jining 272000, Shandong, China Z.H Wang 7900319475328460025219 7900319475328460025219 No Department of Urology, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan 250014, Shandong, China N. Guo 7900319475328460025220 7900319475328460025220 No Department of Urology, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan 250014, Shandong, China J. Tian 198862000666@email.sdu.edu.cn 7900319475328460025221 7900319475328460025221 Yes Department of Urology, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan 250014, Shandong, China