Effects Of Silencing Expression Of Long-chain Non-coding RNA HULC On Proliferation And Invasion Of Human Glioblastoma Cells

Background and purpose:Glioblastoma?GBM?is the most common primary malignant tumor in the brain.Its own strong proliferative capacity and invasiveness are the main obstacles to its successful treatment.At present,the clinical use of surgery,radiotherapy,chemotherapy and biological therapy and other comprehensive treatment,but the patient's survival is not improved,the prognosis is still very poor,the median survival period is about 14months^[1].Current research suggests that the search for a specific treatment regimen for the proliferation and invasion of glioblastoma will likely curb tumor growth and prolong survival.Recent studies have found that long-chain non-coding RNA?LncRNA?has significant expression changes in a variety of tumors,involved in tumor cell proliferation,apoptosis,invasion and metastasis,autophagy and chemosensitivity.The regulation of biological behavior plays an important role in the development of tumors,and may become a molecular marker for invasion and prognosis of GBM,which will provide a basis for targeted therapy of clinical tumors^[2].Highly up-regulated in 1iver cancer?HULC?is a type of LncRNA that has been found to be up-regulated in many malignant tumors,but there are few studies between it and GBM,and the mechanism of action is still unclear.It is well known that a variety of proteins are involved in a series of complex behavioral processes such as tumor cell proliferation and apoptosis,and PI3K/AKT signaling pathway plays an important role in cancer cell proliferation,apoptosis and invasion^[3,4].Combined with the bioinformatics prediction of our group,HULC may play a biological role by activating Focal Adhesion Kinase?FAK?to regulate the expression of upstream and downstream target proteins in the PI3K-AKT signaling pathway,affecting the biological behaviors such as proliferation and invasion of glioblastoma cells.Therefore,in this study,glioblastoma cells SHG44 and U87 cell lines were used as research objects.We tried to iexplore whether LncRNA HULC could activate FAK to regulate PI3K-AKT signaling pathway and promotes the proliferation of glioblastoma cells through cell function and molecular mechanism experiments.Methods:1.Culture human glioblastoma cells SHG44 and U87,and obtain stable cell lines of HULC silent expression group and negative control group by HULC lentiviral interference vector construction technology;2.The proliferation and apoptosis ability of the two groups of cells were detected by CCK8 cell proliferation assay,plate colony formation assay,cell cycle and apoptosis assay;3.The migration and invasion ability of the two groups of cells were detected by cell scratch test and Transwell chamber invasion test;4.The expression level of HULC in the two groups was detected by qRT-PCR.The expression levels of upstream and downstream target proteins?FAK?PI3K?AKT?PTEN?in PI3K-AKT signaling pathway were detected by Western blot.Results:1.The results of proliferation and apoptosis assays showed that the proliferation rate of HULC silent expression group was lower,the colony formation rate was less,the early apoptosis rate was higher,the cells in G1 phase were more,and the number of cells in S phase was less,suggesting that HULC promoted cell proliferation.And inhibit its apoptosis;2.The results of invasion and migration assays showed that the number of cells perforated in HULC silencing group was less and the migration rate was slower,suggesting that HULC promoted cell invasion and migration;3.qRT-PCR results showed that the expression level of HULC in the silent expression group was significantly decreased,that is,the HULC silencing expression stable cell line was successfully constructed;4.The results of Western blot showed that the expression of FAK,PI3K and AKT protein was significantly decreased,and the expression of PTEN protein was higher,suggesting that HULC can exert biological functions by regulating the expression level of upstream and downstream target proteins in PI3K-AKT signaling pathway.Conclusion:LncRNA HULC may promote the proliferation and invasion of glioblastoma cells through FAK activation of PI3K-AKT signaling pathway.