Clinical And Gene Expression Profiles Of Chemoradiotherapy For Localadvanced Squamous Cervical Carcinoma

Objective:At present,about 30%of patients with advanced cervical squamous cell carcinoma are still uncontrolled or recurrenced undergoing standard radical concurrent chemoradiotherapy.This study was to investigate the difference of sensitivity and survival rate of chemoradiotherapy in advanced cervical squamous cell carcinoma,the clinical factors affecting the differernce of sensitivity of chemoradiotherapy,and searching possible related key genes using bioinformatics.Methods:1.Retrospective analysis of the first diagnosis of stage IIB-IVa of locally advanced cervical squamous cell carcinoma from January 2007 to December 2015 in the Department of Gynecology and Oncology,Tumor Hospital Affiliated to Guangxi Medical University and the Department of Oncology,Affiliated Hospital of Southwest Medical University.They were divided into radiochemotherapy sensitive group and radiochemotherapy resistant group,and the survival curve and survival rate were drawn by Kaplan-Meier method using R language package.Log-rank test was used to analyze the difference in survival rate among different groups,and COX regression models were used to analyze the clinical factors affecting prognosis including age,tumor size,pre-treatment hemoglobin level,NLR,PLR,lymph node metastasis status,total radiotherapy time,concurrent chemotherapy course,and adjuvant chemotherapy course.2.Using meta-analysis to analyze the sensitivity of radiochemotherapy in the treatment of advanced cervical squamous cell carcinoma from January 1997to June 2018 and a meta-analysis of this study.After the evaluation of the quality of the literature,the review manager software was used to analyze the heterogeneity of the included documents,and the results of the total effect were obtained.Meanwhile,the published bias funnel diagram was analyzed for the included documents.3.In physical dosimetry,we use the new technique of 3-dimensional computed tomography-guided interstitial brachytherapy,to compare with the traditional three-tube intracavitary brachytherapy.Dosimetric outcomes of the two brachytherapy,incluing the target volume and the dose D90 and the OAR dose.Using SPSS software about Independent sample t test and variance analysis were used to compare the results.4.From the gene expression comprehensive database GEO,two sets of data comparing the radiotherapy sensitive and radiotherapy resistance differential genes were screened,and the differential expression gene was screened using the limma package.The differential genes were screened using the DAVID method for gene pathway enrichment.The pubmed and CCDB databases were used for text mining,and common genes were found.The found genes were verified in OncoLnc for clinical survival data.5.Using the retrospectively clinical patients pathological specimens,the Envison immunohistochemical staining is used to test exptession of the gene which get from bioinformatics.Statidtical treatment by R language Kaplan Meier test,the log-rank test,chi-square test and COX regression model analysis of gene expression and radiation and chemotherapy sensitivity,as well as the impact on the survival and prognostic factors analysis.Results:1.Survival analysis of 965 patients showed that the 5-year survival rate of the radiotherapy-sensitive group was about 40%higher than that of the radiotherapy-resistance group.Tumor staging,hemoglobin level,NLR,lymph node metastasis,and total radiotherapy time were different between the two groups,P<0.01 was statistically significant,However,there was no statistical difference in tumor diameter,PLR,concurrent chemotherapy,and adjuvant chemotherapy.Prognostic analysis of all advanced cervical squamous cell carcinoma:Univariate analysis showed that radiochemotherapy sensitivity,tumor diameter,lymph node metastasis,hemoglobin levels,NLR,PLR,and total radiotherapy time were OS and PFS were prognostic factors.Multivariate analysis showed that radiochemotherapy sensitivity,age,PLR,Radiotherapy time>8 weeks were independent prognostic factors for OS and PFS2.From the published Chinese and English literatures were searched through the Internet search method.Only 7 literatures in Chinese were screened one by one and the results in chapter 1 of this study were included in the meta-analysis.Finally,the literature on the influence of age factors showed good homogeneity and no significant bias,but the total effect showed no statistical difference in radiochemotherapy sensitivity.In the tumor staging,simultaneous chemotherapy treatment and anemia,it is also an unfavorable factor affecting the sensitivity of radiochemotherapy.The included literature has low heterogeneity and less publication bias.In the total radiotherapy time and lymph node metastasis factors,although the total effect suggestion is a disadvantage,after adding our research data,the difference is significantly increased,but the funnel chart is more symmetrical.All studies in the tumor diameter removed our study and another document that caused a large difference,and the literature included was less differential,suggesting that a tumor diameter greater than 4cm was a disadvantage.3.93 patients included in the study,the proportion of chemoradiotherapy resistance was 36.17%and 26.08%,although the volume of the target in the chemoradiotherapy sensitive group was smaller than and there was no significant difference in the dose D90 of the target.The mean volume of HR-CTV and IR-CTV in the interstitial brachytherapy group were 10cm³ bigger than in the intracavitary brachytherapy group.The target dose D90 of HR-CTV and IR-CTV were 101.27 cGy and 60.05 cGy bigger than in the intracavitary brachytherapy group,respectively.The dose of bladder D_0.1cc.1cc was 31.97cGy higher in the interstitial brachytherapy group,but the dose of rectum D_0.1cc,sigmoid D_2cccc and small intestine D_0.1cc.1cc was 47.69cGy,48.92cGy and 40.56cGy,respectively.The organ at risk dose was divided by HR-CTV D90 dose,to normalize.The results showed that there were significant differences in the two groups,except for the sigmoid D_0.1cc.4.From GEO databasese to get 2 sets of data,team need get to 298differentially expressed genes,displayed on the gene function classification gathered mainly in alternative splicing,and splicing variants,AMP rely on synthetic/ligase,acetylation and ATP these five pathways,found in protein interaction between two major ELAVL1 and HSP90AA1 genes,with radiation and chemotherapy and prognosis of cervical cancer occur in the process of database consistency analysis,find seven genes associated with prognosis of cervical cancer and associated with cervical cancer occur in the process of nine genes.In the clinical survival verification with OncoLnc,ASPH and NKAPP1genes were found to affect both radiotherapy sensitivity and survival.5.In the chemoradiotherapy resistance group,ASPH high expression was69.76%;in the chemoradiotherapy sensitivity group,ASPH high expression was40.9%;the mortality rate of 54.2%in the ASPH high expression group was significantly higher than 28.2%in the ASPH low expression group;OS and PFS were significantly reduced in the ASPH high expression group.Univariate analysis showed that high ASPH expression was a prognostic indicator affecting OS and PFS,but multivariate analysis showed no statistical difference.Conclusions:1.A retrospective analysis of 9 years of 965 patients with advanced squamous cell carcinoma of the squamous cell carcinoma showed a significant difference in the 5-year survival rate between the radiation-sensitive group and the resistance group.The main clinical factors that cause this difference are:Tumor stage,hemoglobin level,NLR,lymph node metastasis,total radiotherapy time factors.Through meta-analysis,it can be seen that although there is a certain heterogeneity between the studies,age is not an influencing factor in affecting the sensitivity of chemoradiotherapy in advanced cervical squamous cell carcinoma,other tumors are>4 cm in diameter and tumor staging.lymph node metastasis,hemoglobin<90g/L,total radiotherapy time>8 weeks,synchronous chemotherapy less than 3 weeks are unfavorable factors to reduce the sensitivity of radiotherapy and chemotherapy.2.Prognostic factors analysis of these patients showed that OS and PFS affecting the efficacy of radiotherapy and chemotherapy for advanced cervical squamous cell carcinoma were associated with multiple clinical factors,but chemoradiotherapy sensitivity,age and PLR vv were independent prognostic factors for OS and PFS.3.Using computed tomography-guided interstitial brachytherapy can treat a larger target and can obtain higher doses in the target and lower doses in the organ at risk.It is a safe,effective and worth promoting technology,but it cannot improve the radiochemotherapy sensitivity in local advanced cervical cancer.4.Bioinformatics analysis found 298 differentially expressed genes that play a major role in alternative splicing and splice variant functional pathways,and found that ELAVL1 and HSP90AA1 genes are involved in protein interactions,affecting chemosensitivity.The two genes,ASPH and NKAPP1,were found to affect both radiosensitivity and the genes that affect cervical cancer.5.The main ASPH protein expressed by ASPH gene was highly expressed in the chemoradiotherapy resistance group,and the high expression of ASPH suggested lower OS and PFS and worse prognosis,however,it cannot be used as a sensitivity index to predict chemoradiotherapy.