Research On The Relations Of Genetic Variant To Radiosensitivity And Clinical Outcomes Of Cervical Cancer

Cervical cancer ranks fourth among the most commonly diagnosed cancers in women worldwide and is the second leading cause of cancer-related deaths.Although systemic screening of cervical cancer precancerous lesions and vaccination of HPV vaccines have made it a preventable cancer in recent years,most of them are already in the advanced stage at the time of diagnosis.Concurrent chemoradiotherapy is the standard treatment for locally advanced cervical cancer,but some patients with cervical cancer show resistance to radiation,which leads to a poor prognosis.Therefore,how to improve the curative effect of cervical cancer radiotherapy and reduce the radioresistance of cervical cancer cells has become the key point for research today.In this study,whole exome sequencing will be used to analyze the sensitivity and prognosis-related genes of cervical cancer radiotherapy.Methods:?1?Sequencing analysis: Screening and collecting information on cervical squamous cell carcinoma patients undergoing radical chemoradiotherapy in Sichuan Cancer Hospitals from 2014 to 2016.Cases that do not recur within 3 years after radical chemoradiotherapy are classified as lately recurred groups?LR?,that is,radiationsensitive groups.Conversely,cases that recur within 3 years after radical chemoradiotherapy are classified as early recurred groups?ER?,that is,non-radiation sensitive.One-to-one matching was performed by age,stage,and tumor size before treatment,32 matched cases were selected for whole exon sequencing,and a series of bioinformatics analysis and prognostic analysis were performed based on the sequencing results to discuss whether there are mutation genes related to radiosensitivity and prognosis.?2?Validation analysis: Based on the results of sequencing,independent samples were used to validate the sequencing results.Immunohistochemical analysis,Sanger sequencing analysis,and real-time PCR are used to verify gene mutation,mRNA expression,and protein expression.Results:?1?Results of genomic DNA sequencing revealed that 5 patients in the LR group had single nucleotide variation?SNV?of TRIM64 gene,the type of variation was chr11:g.89701742₈9701743delinsA,and the mutation frequencies were 7.93%,14.38%,8.18%,16.15%,16.07%.However,there were not mutations of TRIM64 gene found in the ER group.In addition,analysis of copy number variation showed that MMP27,DPCR1,YAP1,and BIRC3 genes all had an increase in copy number in the ER group,and there were significant differences from the LR group.The results of RNA sequencing revealed that there were 1072 differentially expressed genes between the ER and LR groups.Among the 1072 differentially expressed genes,the expression level of 7 genes is related to PFS,which are: RP11-335O4.1,CTD-2292M14.1,RP11-94H6.1,RP11-417L19.2,RP4-683M8.2,RP11-20B24.3 and LINC00858.?2?TRIM64 was expressed in the cytoplasm and intercellular substance of cervical cancer tissues through immunohistochemistry.Unfortunately,Sanger sequencing failed to detect mutations of the TRIM64 gene in independent samples.Conclusion: This study found and confirmed for the first time that TRIM64 was expressed in cervical cancer,and there was a significant difference in expression between ER and LR groups.It is suggested that TRIM64 gene mutation may increase the radiosensitivity of cervical cancer tissues.This study also confirmed the correlation between LncRNA and the prognosis of cervical cancer on the basis of other studies,suggesting that the study of cervical cancer at the genetic level is helpful to further understand the radiosensitivity and prognostic factors of cervical cancer.