EGFR Mutations And The High Expression OF GST-Ï€ In Nonsmall Cell Lung Cancer With Clinical Parameters And Prognosis

Objective:To detect the mutation of EGFR gene and the expression of GST-π in NSCLC.To analysis the mutation of EGFR and the expression of GST-π with non-small cell lung cancer in the relationship between clinical pathological features and prognosis.To investigate the mutation EGFR,expression of GST-π and smoking influences in non-small cell lung cancer.Methods:(1)106cases of NSCLC fresh specimens were collected in the second affiliated hospital of nanchang university from2010to2012. While selecting from the tumor edge>5cm of tissue adjacent to carcinoma106cases as control,None of these patients was treated by chemotherapy or radiotherapy before surgery.(2)Using ElivisionTMplus immunohistochemical technology to detect EGFR protein and GST-π protein expression in non-small cell lung cancer; Using real-time fluorescence quantitative PCR to detect EGFR mRNA and GST-π mRNA in non-small cell lung cancer; Using direct sequencing method to detect19and21EGFR gene exon mutations in non-small cell lung cancer.(3)Stafistical analysis was performed using SPSS statistical software package (13.0).Results:1. Immunohistochemical results:(1)EGFR protein1) EGFR protein positive rate was35.8%in non-small cell lung cancer, EGFR protein positive rate was6.6%in tissue adjacent to carcinoma,there have a significant statistical significance difference between the groups(P=0.000);2)Positive rate of EGFR protein in non-small cell lung cancer:①Age<60patients was40.0%,≧60patients was32.8%,there was no statistically significant difference between groups(P=0.444);②Male patients was29.6%,Female patients was56.0%,the difference between groups was statistically significant(P=0.016);③Smoking patients was23.4%,nonsmoking patients was 54.8%,the differences between groups have significant statistical significance (P=0.001);④Squamous carcinoma was19.2%,Adenocarcinoma patients was60.0%, adenosquamous carcinoma patients was37.5%,Large cell carcinoma patients was16.7%,differences between groups have significant statistical significance(P=0.001);⑤Patients with lymph node metastasis was48.1%,Without lymph node metastasis patients was31.6%,there was no statistically significant difference between groups (P=0.096);⑥High degree of differentiation patients was46.4%,Low degree of differentiation patients was16.2%,the differences between groups have significant statistical significance(P=0.002);⑦Clinical stages Ⅰ+Ⅱ patients was43.1%,Stage Ⅲ patients was27.1%,there was no statistically significant difference between groups (P=0.087);(2)GST-π protein1) GST-π protein positive rate was42.5%in non-small cell lung cancer,GST-π protein positive rate was10.4%in tissue adjacent to carcinoma,there have a significant statistical significance difference between the groups(P<0.01);2) Positive rate of GST-π protein in non-small cell lung cancer:①Age<60patients was47.5%,≥60patients was35.6%,there was no statistically significant difference between groups (P=0.217);②Male patients was33.3%,Female patients was72.0%,the difference between groups was statistically significant ((P=0.001);③Smoking patients was39.1%,nonsmoking patients was47.6%,the differences between groups have significant statistical significance (P=0.038);④Squamous carcinoma was30.8%,Adenocarcinoma patients was62.5%,adenosqua-mous carcinoma patients was57.5%,Large cell carcinoma patients was16.7%, differences between groups have significant statistical significance (P=0.023);⑤Patients with lymph node metastasis was55.6%,Without lymph node metastasis patients was37.9%,there was no statistically significant difference between groups (P=0.111);⑥High degree of differentiation patients was34.8%,Low degree of differentiation patients was56.8%,the differences between groups have significant statistical significance(P=0.029);⑦Clinical stages Ⅰ+Ⅱ patients was44.8%,Stage Ⅲ patients was39.6%,there was no statistically significant difference between groups (P=0.587);(3)Mutation EGFR protein and GST-π protein in106cases of NSCLC of Speannan coefficient of rank correlation of r=0.194,P<0.05, correlation analysis showed that EGFR protein with GST-π protein expression were positively correlated in non-small cell lung cancer.2. Real-time fluorescent quantitative PCR results:(1)EGFR mRNA1) EGFR mRNA expression in non-small cell lung cancer was8.33(18.62-10.29),tissue adjacent to carcinoma expression was5.59(15.67-10.08),the differences between groups have statistical significance(P<0.05);2) EGFR mRNA expression in non-small cell lung cancer:①Aged<60patients was4.26±0.27,≥60patients was3.95±0.43,there was no statistically significant difference between groups(P=0.523);②Male patients expression was4.14±0.23,Female patients was5.72±0.34,the difference between groups was statistically significant(P=0.022);③Smoking patients was4.96±0.26,nonsmoking patients was3.95±0.43,the difference between groups was statistically significant(P=0.025);④Squamous carcinoma was4.05±0.52, adenocarcinoma patients was4.13±0.32,adenosquamous carcinoma patients was4.23±0.44, large cell carcinoma patients was4.26±0.27, differences between groups have significant statistical significance(P=0.003);⑤Patients with lymph node metastasis was2.85±0.25, Without lymph node metastasis patients was1.93±0.04, there was no statistically significant difference between groups(P=0.296);⑥High degree of differentiation of patients was2.96±0.44, Low degree of differentiation of patients was4.36±1.83,the differences between groups have significant statistical significance(P=0.004);⑦Clinical stages Ⅰ+Ⅱ patients was4.15±2.03,stages Ⅲ patients was4.14±2.09,there was no statistically significant difference between groups (P=0.407);(2)GST-Ｘ mRNA1)GST-Ｘ mRNA expression in non-small cell lung cancer was11.12(21.41-10.29),adjacent to carcinoma experssion was8.16(18.24-10.08),the differences between groups have significant statistical significance (P<0.01).2)GST-π mRNA expression in non-small cell lung cancer:①Aged<60patients was3.61±0.21,aged≧60patients was3.91±0.13, there was no statistically significant difference between groups (P=0.748);②Male patients was4.14±0.23, Female patients was2.74±0.19,the differences between groups was statistically significant (P=0.041);③Smoking patients was4.32±0.27, nonsmoking patients was3.75±0.23,the differences between groups have significant statistical significance (P=0.037);④Squamous carcinoma was4.65±0.32, Adenocarcinoma patients was 4.33±0.14,adenosquamous carcinoma patients was3.83±0.44,Large cell carcinoma patients was2.26±0.23,differences between groups have significant statistical significance(P=0.008);⑤Patients with lymph node metastasis was2.65±0.35, Without lymph node metastasis patients was3.93±0.74,there was no statistically significant difference between groups (P=0.296);⑥High degree of differentiation patients was4.96±0.27,Low degree of differentiation patients was3.26±1.73,the differences between groups have significant statistical significance(P=0.014);⑦Clinical stages Ⅰ+Ⅱ patients was2.15±0.03,StageⅢpatients was3.94±1.09,there was no statistically significant difference between groups (P=0.757);(3)Content of EGFR mRNA in non-small cell lung cancer is3.12times of tissue adjacent to carcinoma(P<0.05),content of GST-π mRNA in non-small cell lung cancer is1.83times of tissue adjacent to carcinoma(P<0.05).3. Direct sequencing results:EGFR gene extron19in non-small cell lung cancer mutations number is22cases(57.9%), extron21mutations number is16cases(42.1%); EGFR gene extron19in adjacent to carcinoma mutations number is3cases(42.9%), extron21mutations number is4cases(57.1%); comparison two sequencing results EGFR gene extron19and21mutations experiment found that there was no significant difference(x2=3.2,p>0.05).All sequencing results consistent with the real-time fluorescent quantitative PCR detection results, the consistency of100%.Concluison:(1)The expression of EGFR was correlated with sex,smoking history,histo-pathological types and differentiated degree,nonsmoking,femal,high degree of differentiation adenocarcinoma of EGFR mutation rate is high, increase susceptibility to EGFR inhibitor-TKI;and have nothing to do with the patient age,Whether or not lymph node metastasis and TNM staging; EGFR mutation is closely related to the prognosis of patients with tumor, and the prognosis factors, can be used as evaluation of prognosis in patients with clinical indicators.(2)The expression of GST-π was correlated with sex,smoking history,histo-pathological types and differentitated degree, nonsmoking,femal,high degree of differentiation adenocarcinoma of GST-π high expression reduce tumor sensitivity to chemotherapeutic drugs; and have nothing to do with the patient age,Whether or not lymph node metastasis and TNM staging; the high expression of GST-π is closely related to the prognosis of patients with tumor, and the prognosis factors, can be used as evaluation of prognosis in patients with clinical indicators.