Clinical Significance And Expression Of HnRNPK, CyclinD1 And P53 In Human Non-Small Cell Lung Cancer

Objective:To investigate the expression of hnRNPK (heterogeneousnuclearriboncleproten, hnRNPK),CyclinD1 and P53 in NSCLC (Non-Small Cell Lung Cancer) and normal lung tissue, and to explore their possible roles in the tumorigenesis of NSCLC or tumor invasion and metastasis. Method:40 cases of NSCLC specimenes and 20 cases of normal lung tissues were subjected to immunohistochemical staining (SP) for hnRNPK, CyclinD1 and P53. The NSCLC tissues were obtained from 40 patients undergoing lung cancer resection at the Department of Surgery, the Affiliated Hospital of Luzhou Medical College. No patient had received chemotherapy or radiation therapy before surgery. Detailed clinicopathological parameters including age, tumor diameter, stage, lymph node metastasis, and degree of differentiation are as following. There were 29 males and 11 females. Their ages ranged from 28 to 73,whose age<60 were 18 and≥60 were 22. There were 26 squamous carcinomas and 14 adenocarcinomas.The specimenes whose tumor diameter> 3cm were 25 compared 15 specimenes whose tumor diameter≤3cm. The metastatic lymph node specimens were 20, while 20 specimens had no nodal metastasis performance. NSCLC tissues that belongde toⅠ-Ⅱstage were 24 and that belonged toⅢ-Ⅳstage were 16. Moderately differentiated and well-differentiated tumors were 30, poorly differentiated tumors were 10. All the specimens were dealt with formalin fixation, paraffin imbedding and section-cutting. After deparaffinage and hydration, we rinsed the sections with distilled water and soaked them in phosphate buffer saline. Then they were subjected to an antigen retrieval procedure by heating. Endogenous peroxidase activity was blocked using 3% H2O2 solution and nonspecific binding sites were blocked by incubating with goat serum. The slides were incubated with appropriate primary antibody (Rabbit anti-hnRNPK Polyclonal Antibody,Mouse anti-P53 Monoclonal Antibody and Mouse anti-CyclinD1 Protein). Wait 2 hours and then wash them with PBS. The second antibody with biotin labeling were dropped to the slides and incubated. Wait 15 minutes and then wash them with PBS. After that, the slides were incubated with S-A/HRP for 15 minutes and then washed with PBS. Immunologic reaction was developed using DAB. The slides were counterstained with hematoxylin, mounted and then observed through microscope. Consider the cells whose cytoplasm or nucleus had brownish yellow matter as positive cells. The hnRNPK positive signal was localized in both cytoplasm and nucleus, but predominantly in nucleus. The P53 positive signal was mainly in nucleus but CyclinD1 was mainly in cytoplasm. They were divided into 4 levels according to proportion of positive cells and colour depth of each slice.Results:(1)The overexpression of hnRNPK,CyclinD1 and P53 in NSCLC are apparently higher than that in normal lung tissues, this has a significance (P<0.01). (2) The expression of hnRNPK and CyclinD1 are concerned with tumor size and lymph node metastasis, they are in cancer with lymph node metastasis and which more than 3cm were higher than in not with lymph node metastasis and litter than 3cm (P<0.05). There wasn't relation with age, gender, tumor differentiation, TNM sub-period, histological types in NSCLC (P>0.05); The expression of P53 gene correlation with histological type,they in squamouscellcarcinoma was higher than adenocarcinoma (P<0.05). There wasn't relation with age, gender, tumor differentiation, TNM sub-period, lymph node metastasis in NSCLC(P>0.05). (3)There was a positive correlation between the expressions of hnRNPK and CyclinD1 (r=0.379, P=0.016, P＜0.05) and between the expressions of CyclinDl and P53 (r=0.075, P=0.029, P＜0.05).But no relationship was found between hnRNPK and P53 overexpression(r=0.115, P=0.479, P>0.05). Conclusion:(1)hnRNPK protein expression increased in NSCLC was significantly higher than normal lung tissue, and similarly, in NSCLC, the expression of CyclinD1 protein and P53 protein are elevated and significantly higher than normal lung tissue, indicating over-expression of hnRNPK protein,CyclinD1 protein and P53 proteins may have participated in non-small cell lung cancer formation. (2)hnRNPK and CyclinD1 overexpression have relation with tumor size and lymph node metastasis, with lymph node metastasis and cancer which more than 3cm were higher than not with lymph node metastasis and the cancer which little than 3cm, but patient age, histological type, histological grade and TNM stage has nothing to do, indicating that they might not only play a role in tumorigenesis, but also with tumor progression, invasion and metastasis. P53 gene expression have relation with histological type only (P<0.05), in squamous cell carcinoma was higher than adenocarcinoma, indicating that they might only play a role in tumorigenesis. (3) hnRNPK protein was positively related to the higher expression of CyclinD1 in lung cancer. But no correlation with P53. (4)CyclinD1 protein in NSCLC tissues was positively related to the higher expression of P53. they might be affected by the same ways to the occurrence and development of lung cancer.