The Investigation On Biological Effect Of Protamine 1in Non-small-cell Lung Carcinoma

Background:Lung cancer is the most common form of the disease,accounting for nearly12.6% of new cases and 17.8% of cancer-related death worldwide.More than one million people die of lung cancer each year.Non-small cell lung cancer has a low5-year survival rate,poor response to chemotherapy,and high mortality.Surgery is the main treatment for early NSCLC.However,most patients have little chance to undergo surgery because they have locally advanced disease or distant metastases at the time of diagnosis.Therefore,it is very necessary to find a specific molecule for early diagnosis and targeted treatment of NSCLC.Protamines1(PRM1),as a newly discovered tumor testicular antigen,was first identified in the serum and tumor cells of patients with chronic lymphatic leukemia(CLL).Studies have found that PRM1 expression levels in serum and tumor tissues of patients with colorectal cancer are elevated.However,up to now,the function and mechanism of PRM1 in NSCLC have not been reported in depth.In this study,the expression level of PRM1 protein in serum of lung cancer patients and PRM-mRNA in tumor tissues were detected,and the biological function of PRM1 in lung cancer cell lines and nude mice was explored,so as to illustrate the prospect of PRM1 as an early diagnosis and treatment of non-small cell lung cancer.Purpose:1.The expression level of PRM1 in serum and tumor tissues of patients with lung cancer was systematically analyzed to clarify the relationship between the change of PRM1 expression and the occurrence and progression of lung cancer2.To investigate the effect of PRM1 on the biological function of non-small cell lung cancer cell linesMethods:1.The tumor tissue samples of lung cancer patients were collected clinically,and the expression level of PRM1 in the tumor tissues of non-small cell lung cancer patients was detected by RT-PCR and immunohistochemistry,and the correlation between PRM1 and tumor size,clinical stage,pathological grade and lymph node metastasis and other clinical indicators was analyzed by statistical methods.2.Serum samples of patients with lung cancer were collected clinically,and PRM1 protein levels in preoperative serum of patients with non-small cell lung cancer were detected by enzyme-linked immunosorbent assay(ELISA),and compared with those in healthy control group;3.The relationship between serum PRM1 protein expression level and clinicopathological characteristics was analyzed.4.A549 cells and NCI-H460 cells were cultured with different concentrations of serum(0%,2.5%,5%,10%,15%,20%)for 48 h.RT-PCR was used to detect the expression level of PRM1-mRNA,and ELISA was used to detect the secretion level of PRM1 protein in cell culture medium.5.PRM1 protein was added into the culture medium of A549 cells,NCI-H1975 cells and NCI-H460 cells for 48 h,and cell proliferation was detected by CCK8 and EDU methods.6.Anti-PRM1 antibody was added into the culture medium of A549 cells,NCI-H1975 cells and NCI-H460 cells for 48 h,and cell proliferation was detected by CCK8 methods.7.siRNA was used to interfere with the expression of PRM1 in NSCLC cell lines to explore the effects of PRM1 on cell proliferation and cell cycle,so as to evaluate the biological effects of PRM1 on NSCLC cell linesResults:1.The results of RT-PCR,immunohistochemistry and ELISA showed that PRM1 was highly expressed in tumor tissues and serum of patients with NSCLC,slightly expressed or not expressed in normal tissues,and low expressed or not expressed in serum of healthy controls.2.The expression level of PRM1 in non-small cell lung cancer cell lines was increased and the level of PRM1 secreted into serum was increased in the nutrition-deficient state.3.PRM1 protein can promote the proliferation of non-small cell lung cancer.Anti-PRM1 antibodies can inhibit this proliferation.4.After the use of siRNA to interfere the expression of PRM1 in NSCLC cell lines,the proliferation of NSCLC cell lines was significantly inhibited.Conclusion:Through this topic research,we first discovered the PRM1 expression in non-small cell lung cancer,and clear the PRM1 correlation with non-small cell lung cancer development,reveals the PRM1 involved in regulating the proliferation of small cell lung cancer,further enrich the study of the pathogenesis of lung cancer and for PRM1 antibody could be a new treatment strategies non-small cell lung cancer.