Expression Of HnRNP L Protein In Human Clear Cell Renal Cell Carcinoma Tissues And Its Clinical Significance

Background and objectiveRenal Cell Carcinoma (kidney), which is originated from malignant tumour of the urinary tubular epithelial cells of Renal parenchyma, is high grade tumor in urinary system, also is one of the most common tumor, also called Renal adenocarcinoma, accounting for 85%of the primary tumor kidney-90%, of which 80%-90% of Clear Cell Carcinoma (Clear Cell takes Cell Carcinoma), and its incidence is rising in recent years.Due to the pathogenesis of renal clear cell carcinoma is unclear, the early clinical manifestation is not typical, often found in nearly half of the patients had end-stage kidney or distant metastasis, and loss of opportunity of operation.Kidney cancer is not sensitive to radiotherapy and chemotherapy are at the same time; Efficacy of immunotherapy is limited, therefore, its most main treatment is surgical resection, but the postoperative recurrence rate of 25%-50%.In general, drugs and surgery for renal cell carcinoma patients progression-free surial and oerall surial remains unsatisfactory.The lack of specific molecular markers and therapeutic targets effectively is the process of diagnosis and treatment of renal cancer and the problems facing the key.Obviously,renal clear cell carcinoma clinical diagnosis way of breakthrough and the improvement of treatment depends on the significant progress of basic research, depends on renal clear cell carcinoma mechanism to clarify.A growing body of evidence suggests that RNA alternative splicing as a widespread biological processes, in the process of the occurrence and development of tumor play an important role.Among them, the uneven nuclear ribonucleoprotein family (hnRNPs) by selective splicing occupy the important position in the tumorigenesis.Heterogeneous nuclear ribonucleoprotein L (hnRNP L) as a member of the of the family of hnRNPs, successively carried out in the tumor research in recent years, but the hnRNP L expression in renal clear cell carcinoma and its function regulation mechanism research at home and abroad, there is no publicly reported.hnRNP L is found in the nucleus of a class of RNA binding protein, so far have found about 20 many hnRNPs (a～U) protein, has found a variety of hnRNPs protein in tumor occurrence and development plays an important role.HnRNP L is found in the early transcription segments of rich cystine and proline ribonucleoprotein and it is a important part of heterogeneous nuclear ribonucleoprotein. HnRNP L as RNA binding protein, participate in chromosome remodeling, regulation and signal transduction process of transcription, translation, in its role in the development of tumor is more and more attention.Existing bioinformatics analysis also showed that hnRNP L and hnRNPs several family members are partners (function partner), analysis Pubgene hnRNP L and cell biological processes, such as apoptosis, death and growth are closely related.In recent years, successively have hnRNP L study reported in the tumor may be affected by precursor mRNA splicing hnRNP L involved in liver cancer, lung cancer and metastatic colorectal cancer and squamous cell carcinoma of the esophagus cancer, but its molecular mechanism and may involve signaling pathway are not yet fully clear.From published literature did not see the role of hnRNP L in renal clear cell carcinoma mechanism research, but hnRNP L mechanism research has been reported in other related fields.It may be through the following mechanisms play a role:(1) Precursor mRNA splicing:hnRNP precursor with caspase-9 L mRNA combining the aggressivity of the regulation of lung cancer cells; At the same time hnRNP L also participate in CEACAM1 precursor mRNA alternative splicing;(2) the IRES (internal ribosome to site sequence) to adjust:IRES of non-coding RNA, folded into a structure similar to the initial tRNA, thus mediated ribosomes and RNA, starting protein translation. With the literature, hnRNP L function depends on the HCV IRES activity;(3) reverse transcription regulation:hnRNP L can be used as a main influence factors by combining the LINE 1 (long dispersed element) RNA in reverse transcription regulation;(4) interact with micrornas:Jafarifar and found that there are four short chains of non-coding RNA (miRNA) in VEGFA (vascular endothelial growth factor) 3’-noncoding OuDeShun effect element, and hnRNP L combination in the same area, and it can inhibits the silence of micrornas.HnRNP L relationship with the occurrence and development of tumor has been clear, however, so far, it happens in renal clear cell carcinoma of the function and its molecular regulation mechanism is still in the development of many important questions to clarify. HnRNP L involved in signaling pathways is still not clear; Clarify hnRNP L gene expression regulation mechanism is clear its role in cancer development important foundation.This research will help find renal cell carcinoma pathogenesis; providing new molecular markers for clinical renal clear cell carcinoma of early diagnosis, treatment and prognosis judgementMethod(1) Taking about the 3mmx5mm size of each cases of renal clear cell carcinoma tissue, drawn tube, immediately after put into cryopreserved in liquid nitrogen. According to the preliminary divided into pathological diagnosis of renal clear cell carcinoma group and normal kidney tissue. Western blot test hnRNP L protein in the renal clear cell carcinoma tissues and the expression of tumor tissue and kidney from has collected 90 specimens renal clear cell carcinoma tissues and its matching in the tumor tissue adjacent to carcinoma hnRNP L protein expression level verification, to further verify the expression level and renal clear cell carcinoma of the correlation between histologic grade. Immunohistochemical detection of two groups of samples hnRNP L expression intensity. Dyeing express grading methods according to the staining intensity and the proportion of positive cells number two indicators for evaluation. Then apply statistical analysis hnRNP L and the correlation between the expression and clinical parameters(Include:age, sex, tumor size, TNM stage, pathological stage, histologic grade, etc).(2) HnRNP L protein expression in renal clear cell carcinoma of the relationship with the prognosis of patients, Kaplan Meier method analysis HnRNP L expression with the relationship between the survival time. In addition, we of hnRNP L detection sensitivity and the degree of specific statistical analysis, and determine whether it can be used as the diagnosis of renal clear cell carcinoma molecular markers.(3) The function of hnRNP L have begun to be studied by other team members, By adopting Arraystar Human LncRNA Microarray V3.0 chip, compared the knock on low hnRNP L kidney SN12-PM6 cells and control cells LncRNA and mRNA gene expression differences, preliminary analysis hnRNP L the possible regulatory mechanism in renal clear cell carcinomaResults(1) HnRNP L protein in 90 cases of renal clear cell carcinoma tissues and there are differences between the expression of the tumor tissue adjacent to carcinoma, for lower expression in tumor.HnRNP L protein expression mainly in the nucleus, diffuse in distribution. HnRNP L protein expression in renal clear cell carcinoma tissues positive rate was 71.1% (64/90); Protein in renal clear cell carcinoma tissues hnRNP L color on the number of positive cells and the color intensity of the non tumor tissue adjacent to carcinoma significantly reduced, in a small number of negative expression in the tumor tissue adjacent to carcinoma, the positive expression rate was 98.9%(89/90), the comparison, statistically significant difference (P< 0.05);(2) 90 cases of renal clear cell carcinoma tissues, hnRNP L protein expression and T stage, organization classification, clinical stage and closely related to tumor size. Among them, Tla, Tl b and T2, T3 hnRNP L protein positive expression rate was 88.9%(32/36),76.7%(23/30) and 37.5%(9/24), comparison between groups, the difference was statistically significant (P< 0.05). Histological grading of Ⅰ, Ⅱ and Ⅲ patients, hnRNP L protein positive expression rate of significant difference (P< 0.05). Clinical staging in patients with stage 1 and 2, hnRNP L protein positive expression rate was 83.1%(54/65) and 40.0%(10/25), the comparison between groups, statistically significant difference (P< 0.05). Tumor size (178 or less,> 178), hnRNP L protein positive expression rate were 82.0%(50/61) and 48.3%(14/29), compare the differences between groups is significant (P< 0.05). However, hnRNP L protein expression has no obvious correlation with the patient age;(3) Kaplan Meier method analysis showed hnRNP L lower expression patients survival time was significantly lower than the high expression of patients, hnRNP L total survival time in patients with low expression (37.75+4.904) is shorter, hnRNP L overall survival in patients with high expression (68.483+3.286) for a long time, the difference is statistically significant (Log Rank=30.18, P=0.000) (bold lines represent hnRNP L low expression group; the dotted line represents the hnRNP L high expression group). HnRNP L positive patients 5-year survival rate was 75.0%(95% CI,64.024%,85.976%), however, HnRNP L negative patients 5-year survival rate is only 20.8%(95% CI,4.532%,37.068%); Survival time in patients with tumor volume less than 178 cm3 data analysis shows that in small size hnRNP L differentially expressed in renal cancer patients survival time also can produce significant differences (P< 0.001);Survival time in patients with histological grade in phase I of the data analysis shows that stage I of hnRNP L differentially expressed in early renal cancer patients survival time also can produce significant differences (P< 0.001); Histologic classification in patients Ⅱ-Ⅲ period also have significant difference (P= 0.007); Patients with clinical stage I also more significant difference (P< 0.001); TNM staging of pT stage for stage I patients also significant difference (P< 0.001);Multiariable Cox risk ratio model analysis showed that:hnRNP L is independent risk factors for the prognosis of patients with renal clear cell carcinoma (P= 0.009).(4) Analyzing differences lncRNA/mRNA in the intersection, in all 33000 lncRNA and 30200 gene probe, the screening of hnRNP L expression level difference lncRNA 4506 (raisedConclusion(1) HnRNP L protein in human renal clear cell carcinoma in the expression level was significantly lower than the tumor tissue adjacent to carcinoma, renal clear cell carcinoma, and negatively correlated with pathological grading, the expression level of worse outcomes for patients with drop hints;(2) HnRNP L can be used as an independent risk factor for the prognosis of patients with renal clear cell carcinoma.(3) HnRNP L is associated with renal clear cell carcinoma(ccRCC) development, its potential as a renal clear cell carcinoma patients with early diagnosis and prognosis evaluation of molecular markers.(4) HnRNP L may through the interaction with lncRNA to participate in the occurrence of renal clear cell carcinoma.