Preliminary Research Of Long Non-coding RNA CUDR’s Function And Mechanism In Pancreatic Ductal Adenocarcinoma

ObjectiveThis project is to to gain the different expressed lncRNAs and mRNAs in PDAC, and to predict the potential and critical ones of those specific lncRNAs by bioinformaticly analyzing the lncRNA/mRNA profiling of surgical specimens between pancreatic ductal adenocarcinoma(PDAC) and para-carcinoma tissue. Then, the expression level of these lncRNAs should be verified in the clinical samples, and be analyzed the relationship with clinical data in cancer’s development. Furthermore, we will detect the influence of the specific lncRNA to the pancreatic cancer cells’ biological behavior and its mechanisms.Methods1、The different expressed lncRNAs and mRNAs between PDAC and tumor-adjacent tissues were obtained by 5 pairs of microarrays, and hierarchical cluster analysis should be done to confirm the differences.2、GO analysis, Pathway analysis and lncRNA-mRNA network were performed to find the potential and critical lncRNAs, and to predict the latent pathways and targets of those specific lncRNAs, along with searching published literatures and public databases on the internet.3、Find one of those potential and critical lncRNAs as further object of study. To investigated the expression level of the chosen lncRNA in clinical samples by quantificational real-time polymerase chain reaction. And the relationship between those clinic data and the lncRNA expression level in pancreatic cancer’s growth, malignant grade, invasion and migration should be analyzed statistically.4、Pancreatic carcinoma cell line expressing the chosen lncRNA at a high level stably should be established. Then to detect the function and mechanisms of CUDR in regulating the biological behavior of pancreatic cancer by cell experiments in vitro, such as MTT, Transwell, wound healing, Cell clone forming, Western Blot, and so on.Results.1、According to the standard, the expression of lncRNA/mRNA in PDAC’s microarrays increased or reduced 5 times and P< 0.01,414 lncRNAs and 713 mRNAs were gained. Then, hierarchical cluster analysis was done, and its result indicated that the different expressed mRNAs and lncRNAs were credible.2、Analyzing bioinformaticly the differential expressed mRNAs and lncRNAs,77 raised pathways and 33 decreased with statistical significance were obtained. Among them, more than ten signaling pathways have been supposed to regulate pancreatic cancer’s development.7 mRNAs (YWHA、CDH3、NTRK1、BIRC3、LAMC2、KRAS、 ITGA2), affiliated to pancreatic cancer-concerned pathways, were in the center of the established coexpression network.11 lncRNAs were highly relative to those 7 mRNAs and also in the central part of the co-expression network.3、Among 11 lncRNAs, CUDR was chosen to further study. Its high expression level in pancreatic cancer issues was confirmed in 20 clinical samples by qRT-PCR. Statistical analysis showed that, the expression level of CUDR had something to do with tumors’ differentiation invasion and lymphatic metastasis.4、We successfully built a pancreatic carcinoma cell line,Panc-1-CUDR, which expresses CUDR at a high level stably and its negative control cell line,Panc-1-MOCK, using total gene synthesis and retroviruses transfection technique. By experiments in vitro, we found that, CUDR could enlarge the ability of Panc-1’s proliferation, migration, invasion and clone formation. Its mechanism probably was to active AKT signaling pathway and to stimulate Panc-1’s epithelial mesenchymal transition (EMT).Conclusions1、lncRNAs are playing an important part in PDAC’s developing. Accompanied further studies, they may become important diagnostic, prognostic or theoretic targets in the future.2、CUDR can stimulate pancreatic carcinoma’s malignant biological behavior, such as proliferation, migration, invasion and clone formation. And its mechanism perhaps was to regulate FAK/PI3K/AKT signaling pathway and to promote pancreatic carcinoma cells’EMT.