| Objective To explore the correlation between estrogen receptor alpha gene(ESR1)polymorphism and the risk of bladder and kidney cancer in Chinese population.This study will identify the relationship between ESR1 gene polymorphism at four loci and the clinicopathological characteristics of bladder and kidney cancer.We also investigate interaction on bladder and kidney cancer between genetic polymorphisms and environmental factors such as living environment and lifestyle.Methods(1)This study used a 1:1 matched case-control study.We recruited newly diagnosed hospitalized patients in the affiliated hospital of Inner Mongolia Medical University and Inner Mongolia Autonomous Region People's Hospital from January 2018 to December 2019.According to the inclusion criteria,we selected 257 patients with bladder cancer and 110 patients with kidney cancer as the case group.The control group was selected from healthy individuals undergoing physical examinations in the same term.The controls were matched 1:1 by gender and age ± 5 years.All subjects were required to sign an informed consent.(2)The basic information is collected by a questionnaire,which includes general conditions,occupational history and past medical history,living habits and environmental conditions,and family medical history.We collected clinical laboratory results such as HGB,RBC,WBC,NE% through blood test report.The pathological type,pathological grade,clinical stage of TNM,and distant metastasis of the tumor were collected by pathological test report.(3)Collection and preservation of blood specimens: In the early morning fasting state(fasting and water-free for 6-8 hours),2 ml each of intravenous EDTA anticoagulant and inert separation gel procoagulant were collected,and serum and blood cells were collected after centrifugation,stored at at-80 ° C freezer.(4)Genotype detection: Extract the blood genomic DNA of the research object,design PCR primers,and respectively amplify the ESR1 gene(rs2077647,rs1801132,rs2234693,rs9340799)loci,and use multiple high temperature ligase detection reaction technology(im LDR)to pair The four loci of ESR1 gene were tested for SNP typing.(5)Enzyme-linked immunosorbent assay(ELISA)to detect ESR? levels in serum: Take 100 ?l serum to detect serum ESR? levels in the case and control group according to the instructions of the human estrogen receptor alpha enzyme-linked immunoassay kit.(6)Statistical processing: All statistical analysis was used IBM SPSS 25.0 statistical software,according to the nature of data and study purpose.If data obey the normal distribution,the measurement data were expressed by "mean ± standard deviation".The non-submission normal distribution were expressed by "median(interquartile range)".The count data were expressed by frequency(%).Comparison of two sets of measurement data such as obeying normal distribution,uniform variance using paired t test,non-normal distribution or uneven variance using rank sum test.Comparison of qualitative data between the two groups using ?2 test.The ?2 test was used to analyze whether the genotype frequency distribution of the four loci of the ESR1 gene accorded with the Hardy-Weinberg(H-W)genetic balance.Multivariate conditional logistic regression was used to calculate the odds ratio(OR)and its 95% confidence interval(CI)to assess the relationship between life behavior,eating habits and genetic polymorphisms and the risk of bladder and kidney cancer relationship.Generalized multi-factor dimensionality reduction method(GMDR)was used to analyze the correlation between ESR1 gene polymorphisms and bladder cancer and kidney cancer,and the gene-environmrnt interaction was analyzed.Results(1)Univariate and multivariate analysis showed that compared with the control group,bladder cancer cases had significant differences in smoking,drinking,dietary tastes,holding back urine,physical activity,cooking with animal oil,fruit intake,and physical exercise(P<0.05),and their OR values were 3.940,2.554,2.079,3.609,22.534,2.440,0.392,0.610,respectively.There were significant differences(P<0.05)between kidney cancer group and control group in many respects,including smoking,dietary supplements,physical labor,and urination,their OR values were 6.286,5.976,13.937,and 5.156,respectively.There were no significant differences in physical exercise,BMI,and fruit intake between the two tumors and their control groups(P> 0.05).(2)Among the four polymorphic sites of the ESR1 gene,the genotype frequencies of the rs1801132 and rs2077647 loci between the bladder cancer case group and the control group were statistically different(P<0.05),and rs1801132 GG/GC may be a protective factor for bladder cancer(P=0.018,OR=0.523;P<0.001,OR=0.481),rs2077647 TT/CT may be a risk factor(P=0.005,OR=2.621;P=0.015,OR= 2.386).There was no significant correlation in rs2234693 and rs93440799 with bladder cancer.However,in the correlation study of renal cancer,the four locus of ESR1 gene were not correlated with the kidney cancer risk.(3)In the risk of bladder cancer,there are potential gene-environment interactions between the rs1801132 and rs2234693 sites of ESR1 gene and smoking,salty diet,lack of exercise,etc.The rs1801132 and rs2077647 loci have potential interactions with physical/mental labor and frequent holding back urine in the risk of kidney cancer.(4)The genotype distribution of rs1801132 was statistically different on patient with bladder cancer in pathological grade,clinical stage,and distant metastasis(P<0.05).The rs2077647 and rs9340799 alleles were distributed clinically.The differences in stages were statistically significant(P<0.05).In renal cancer,the rs1801132 allele frequency was statistically different in pathological type and clinical stage(P<0.05),and the rs9340799 genotype distribution was statistically significant in clinical stage and distant metastasis(P<0.05).There was no significant difference in rs2234693 genotype and allele frequency distribution between clinical indicators of the two tumors(P>0.05).(5)In the clinical examination data of bladder cancer,HGB,RBC,WBC,NE% were statistically different between the two(P<0.05),and the HGB and RBC indicators of the case group were lower than those of the control group.The comparison of the two groups was(141.08±22.44g/L,145.86±19.51g/L)and [(4.57±0.72)×10^12/L,(4.77±0.61)×10^12/L],while WBC and NE% were higher than the control group,they were [(7.37±3.73)×10^9/L,(6.76±2.30)×10^9/L] and(61.97±11.97,59.73±11.52),respectively.The differences of the four haematological indexes between the renal cancer case group and the control group were also statistically significant(P<0.05),and the HGB and RBC of the case group were significantly lower than those of the control group.The two groups were(137.01±20.85g/L,145.24±22.76g/L)and[(4.56±0.63)×10^12/L,(4.76±0.68)×10^12/L],while WBC and NE% were significantly higher than those in the control group,which were [(7.54±2.52)×10^9/L,(6.82±2.55)×10^9/L] and(65.17±11.93,61.53±13.21).(6)The serum ESR? protein concentration was detected by ELISA.The expression of ESR? protein in the serum of bladder cancer cases was significantly lower than that of the control group(51.70±12.21pg/ml,64.22±13.46pg/ml),and the difference was statistically significant(t=13.394,P<0.001).There was no significant difference in the expression of ESR? protein between the renal cancer case group and the control group(t=0.489,P=0.628).Conclusion(1)Smoking,drinking,salty diet,urination,and overwork may be risk factors for bladder cancer.Fruit intake and physical exercise are protective factors for bladder cancer.Smoking,eating too much meat,overwork,and urination may be a risk factor for kidney cancer.(2)Our results show that the ESR1 gene rs1801132 GG/GC may be a protective genotype of bladder cancer,and rs2077647 TT/CT may be a risk genotype for bladder cancer.(3)In the risk of bladder cancer,there are potential gene-environment interactions between the rs1801132 and rs2234693 loci of the ESR1 gene and environmental factors,such as smoking,salty diet,and lack of exercise.The rs1801132 and rs2077647 loci have potential interactions with physical/mental labor and frequent holding back urine in the risk of kidney cancer.(4)In bladder cancer patients,the genotype distribution at rs1801132 was statistically related to pathological grade,clinical stage,and distant metastasis,and the allele distribution at rs2077647 was statistically related to the clinical stage.In renal cancer,the allele frequency of rs1801132 locus is statistically related to pathological type and clinical stage,and the genotype distribution of rs9340799 locus is statistically related to clinical stage and distant metastasis.(5)The contents of HGB and RBC in the bladder cancer and kidney cancer cases were lower than those in the healthy control group,and the WBC content and NE% were higher than those in the control group.(6)The expression of ESR? protein in bladder cancer cases was significantly lower than that in the control group. |
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