The Relationship And Expression Of WWOX, STAT3 And C-myc In Human Non Small Cell Lung Cancer

Abstract:Objective:To investigate the expression of STAT3,WWOX and c-myc in non small cell lung cancer(NSCLC) and their clinical significance,and study their relationship. Method: Immunohistochemistry SP method to detect WWOX, STAT3, c-myc in 40 cases of non-small cell lung cancer and 20 normal lung tissues, and analyze its relationship with clinicopathological parameters. NSCLC, surgical resection were obtained without any preoperative radiotherapy or chemotherapy.29 cases were male and 11 female, aged 35 to 78 years, of which<60 years of age in 22 patients,≥60 years of age in 18 cases. According to WHO histological classification, staging,26 patients with squamous cell carcinoma, adenocarcinoma 14; including lymph node metastasis in 18 cases,22 cases without lymph node metastasis; tumor≤3 cm in 14 cases,> 3 cm 26 cases; 30 cases of high grade, poorly differentiated 10 cases,Ⅰ-Ⅱ,24 cases,Ⅲ-Ⅳof 16 cases. The specimen tissue fixation, dehydration, transparent, made of tissue after paraffin wax block, conventional 4μm serial sections. After routine dewaxing sections hydration, distilled water, phosphate buffer (phosphate buffer saline, PBS) immersion, high temperature antigen retrieval,3%H2O2 block endogenous peroxidase activity, and closed with normal goat serum cross-reaction. Dropping an anti-(STAT3 rabbit anti-human polyclonal antibody, mouse anti-human WWOX monoclonal antibody, mouse anti-human c-myc monoclonal antibody) incubation, PBS washed dropping incubated with biotinylated secondary antibodies, PBS washed dropping horseradish enzyme labeled streptavidin working fluid (SA/ HRP) incubation, PBS washing, DAB color, hematoxylin, transparent, Fengpian observation. To occur within the cytoplasm or nucleus of brown particles, shape or brown reticulated positive cell clumps. Which WWOX protein mainly in the cytoplasm, the expression of positive signals of STAT3 protein in NSCLC tissue mainly in the cytoplasm, a few in the nucleus, the expression of c-myc protein was mainly found in the cytoplasm; Three caught by semi-quantitative integration method to judge the results, the observation of each section 10 high power field, cells per high power field in mind the percentage rate and color depth for:(1) positive cells according to the percentage of positive cells Rating:≤5% positive cells was 0 score,6%to 25%for 1 score,26%to 50%for 2 score, 51%to 75%for 3 score,> 75%compared with 4 score, (2) according to color intensity score:no color is 0 points,1 point, yellow, brown for 2 score, brown for 3 score. Access (1) (2) of the score and value as the yardstick staining:0 into negative (-),1-2 divided into weak positive (+), positive (++),3 to 5 were divided into 67 is divided into strongly positive (+++) expression. Results:①The expression of WWOX in non-small cell lung cancer is 30%, significantly lower than normal lung tissue (75%);The positive expression of STAT3 in non-small cell lung cancer was 92.5%, significantly higher than the normal lung tissue (20%);The positive expression of c-myc in non-small cell lung cancer is 90%,significantly higher than the normal lung tissue (15%). After statistical analysis, they are in tumor tissue and normal lung tissues were statistically significant difference (P<0.01).②The expression of WWOX wasn't correlated with age, gender, TNM stages, tumor size and tumor differentiation, which is ralated to whether lymph node metastasis in NSCLC(P<0.01), with lymph node metastasis non-small cell lung cancer loss of WWOX gene expression or lower expression.The levels of STAT3 and c-myc were relation with histological types in non-small cell lung cancer, in adenocarcinoma was significantly higher than squamous cell carcinoma (P<0.01), The overexpression of STAT3 and c-myc weren't associated with age, gender, tumor differentiation, TNM stages, tumor size and lymph node metastasis (P>0.05).③The levels of STAT3 protein and WWOX in non-small cell lung cancer was negatively correlated (r=-0.443, P<0.01);④There was no correlation between the expressions of STAT3 and c-myc (r=-0.084; P>0.05);⑤The expression of WWOX and c-myc protein in non-small cell lung cancer had no correlation (r=0.115, P>0.05). Conclusion:①STAT3 protein expression increased in NSCLC was significantly higher than normal lung tissue, and similarly, in NSCLC, the expression of c-myc protein is elevated and significantly higher than normal lung tissue, indicating over-expression of STAT3 protein and c-myc proteins may have participated in non-small cell lung cancer formation. WWOX protein in NSCLC tissue was low.②The expression of WWOX gene was related to whether the lymph node metastasis, lymph node metastasis of lung cancer in the tissues decreased, loss or reduction of its expression maybe related to with tumor aggressiveness. The level of STAT3, c-myc were correlated with histological types in non-small cell lung cancer, which in adenocarcinoma was significantly higher than squamous cell carcinoma, and nothing to do with patient age, tumor size, histological grade, lymph node metastasis and TNM stage, indicating that they might only play a role in tumorigenesis, but wrern't related with tumor progression, invasion and metastasis.③There was negatively related between expression STAT3 protein and WWOX protein,the higher expression of STAT3 in lung cancer, WWOX expression in the less or were absent,which may be indictive of their antagonism.④There was no correlation between expression of STAT3 and c-myc protein in NSCLC, indicating the formation and development of tumors is induced by c-myc pathway,which was not through SATT3.⑤c-myc and the expression of WWOX was no correlation, they may be affected by two different ways to the occurrence and development of lung cancer.