The Expression And Mechanism Of Long Non-coding RNA HOXC-AS1 In Glioma

Glioma is the most common primary malignant tumors in the adult central nervous system,accounting for more than 40% of intracranial tumors,and the proportion of high-grade gliomas is high.The current treatment of glioma to surgery,assisted with radiotherapy and chemotherapy.Although in recent years the treatment of glioma has made some progress,the prognosis of patients with malignant glioma did not achieve the desired effect,malignant gliomas are still seen as one of the tumors that have not made significant progress in the treatment of tumors.There is no doubt that the development of tumors is a multifactorial process.With the deepening of understanding of the tumor,people began to study from the genetic level of tumor development.Therefore,to explore the occurrence and development of glioma molecular mechanism,become the current hot spots of glioma research.Long non-coding RNA(lnc RNA)is a class of transcripts that are more than 200 nucleotides and with no function of encoding protein.At first,it is considered to be "noise" in gene transcription and does not have biological function.However,recent studies have shown that long non-coding RNA plays an important role in the regulation of gene expression and epigenetics,and can participate in the regulation of cell differentiation and individual development at multiple levels.Although long non-coding RNA investigations of the tumor have obtained some achievements,the vast majority of lnc RNA mechanism is still unclear.Thus,this is also an important direction for future biological research.We have used a microarray data to find a long non-coding RNA HOXC-AS1 that is abnormally expressed in gliomas,and there is no report on cancer research,so it is also our interest.This study first clarified the expression of long non-coding RNA HOXC-AS1 in glioma tissue and normal brain tissue,combined with the clinical indicators and prognosis of patients,the statistical analysis of its relevance.According to genetically engineered interference target HOXC-AS1,and study glioma cell changes of biological behavior by using in vitro functional experiments.We want to find the possible target gene HOXC8,and to explore preliminary mechanism of HOXC-AS1.Our study was consisted of three parts.The first part is to detect the expression of HOXC-AS1 in glioma tissues.The second part is to investigate the biological behavior of glioma cells U251 and A172 after down-regulation of HOXC-AS1.The third part is to screen the target gene HOXC8 of HOXC-AS1 and to study preliminary mechanism.Part ? the expression of lnc RNA HOXC-AS1 in gliomaObjective: To investigate the relative expression of long non-coding RNA HOXC-AS1 in glioma and normal brain tissue.To analyses the correlation of relative expression of HOXC-AS1 with clinical indicator of patients.Methods: The rerelative expression leavel of HOXC-AS1 in diferent grades glioma and normal brain tissues was detected by real-time quantitative PCR.The correlation between the expression level of HOXC-AS1 and various clinical indicators were analyzed by statistics.And the correlation between the relative expression level of HOXC-AS1 and the prognosis of glioma patients was analyzed by Kaplan-Meier survival.Results: The relative expression of HOXC-AS1 in different pathological grade gliomas and normal brain tissues was detected by q RT-PCR.The results showed that the relative expression of HOXC-AS1 in high-grade glioma tissues was significantly higher than that in low-grade glioma tissues and normal brain tissues.Combined with various clinical indicators of patients,the results showed that HOXC-AS1 expression was positively correlated with glioma pathological grade.At the same time,the relative expression level of HOXC-AS1 was negatively correlated with the overall survival time of patients by Kaplan-Meier survival analysis.Conclusion: The relative expression level of HOXC-AS1 in different glioma specimens was positively correlated with glioma pathological grade.The relative expression level of HOXC-AS1 was negatively correlated with the prognosis of glioma patients.Part ? the effects of down-regulation of HOXC-AS1 on the biological behavior of glioma cells U251 and A172Objective: To study the functional role of HOXC-AS1 in glioma cell linesMethods: A series of in vitro functional changes of glioma cells after knockdown of HOXC-AS1 were investigated by Lipofectamine 3000 transfection of glioma cell lines.Results: After transfection of Smart Silencer into glioma cell lines U251 and A172,q RT-PCR results showed a significant decrease in HOXC-AS1 expression.The proliferation of glioma cells was decreased after knockdown of HOXC-AS1.Transwell experiments showed that the migration and invasion of glioma cells decreased after knockdown of HOXC-AS1.The results also showed that the apoptotic proportion of glioma cells increased after knockdown of HOXC-AS1,and the cell cycle and cycle-related proteins were changed.Conclusion: HOXC-AS1 can promote the proliferation,invasion and migration and inhibit the apoptosis of glioma cells.Part ? the study of HOXC-AS1 target gene HOXC8Objective: To study the correlation of HOXC-AS1 and HOXC8 in glioma tissue samples.To explore the possible mechanism of HOXC-AS1 in glioma.Methods: Bioinformatics analysis was used to identify the possible target genes of HOXC-AS1.The expression level of HOXC8 in different glioma tissues and normal brain tissues was detected by using real-time quantitative PCR.Pearson's correlation analysis was used to detect the relationship between the expression of HOXC-AS1 and HOXC8 in glioma tissues.The expression of HOXC8 in tissue samples was detected by Western blot and immunohistochemistry.The changes of HOXC8 at the protein level after knock-down of HOXC-AS1 were detected by Western blot in glioma cell lines.Results: The results of q RT-PCR showed that HOXC8 was decreased when down-regulation of HOXC-AS1.Pearson's correlation analysis showed that HOXC-AS1 and HOXC8 were positively correlated in different glioma samples.Western blot and immunohistochemistry showed that HOXC8 expression level was positively correlated with glioma pathological grade.The results of Western blot also showed that knockdown of HOXC-AS1 could inhibit the expression of HOXC8.Conclusion: HOXC-AS1 may play a role in carcinogenesis in glioma cells by regulating the expression of HOXC8.