The Study On The Role And Mechanism Of LSM14A In Glioma

Objective: Primary CNS tumors are a heterogeneous group of tumors arising from cells within the central nervous system.Gliomas are the most common malignant primary brain tumors,accounting for 81% of malignant brain tumors.Although its incidence is relatively rare,patients with glioma tend to have high mortality and recurrence rates,with their 5-year overall survival rate not exceeding 35%.Adult diffuse infiltrative gliomas are now divided into three overall groups with different natural course,treatment response and prognostic outcome: tumors with isocitrate dehydrogenase(IDH)mutations and 1p/19 q co-deletions,which are predominantly oligodendrocyte morphology,have the best prognosis;tumors with IDH mutations and 1p/19 q non-co deletions,which are predominantly astrocytic in histology,have an intermediate prognosis;IDH wild type,predominantly higher WHO grade(III or IV)tumors,have a poor prognosis.Therapeutic studies of glioma have shown that a series of growth-regulating related molecules such as growth factors and cytokines play an important role in glioma growth,invasion,and treatment tolerance.Therefore,the regulation of glioma-related molecules has become an important target for glioma therapeutic research.The like-Sm(LSm)protein family has more than a dozen members that are highly conserved across species and are involved in various signaling pathways related to RNA metabolism.Members of this family form loop complexes on specific target RNAs and play key roles in their biosynthesis,function and translocation.lsm14A(LSM14A m RNA processing body assembly factor)is one of the LSM protein family members in the two heptameric loop structures formed by the LSM1-8 proteins.LSM14 A can function as an alternative structure to both complexes.It is also an essential component in the formation of m RNA processing bodies(P-bodies)and stress granules(SGs),which are involved in the translational repression of m RNAs and provide storage sites for untranslated m RNAs.It has been shown that P-bodies and SGs promote tumor proliferation and development by regulating several important typical signaling pathways,such as mammalian target of rapamycin(m TOR)and mitogen-activated protein kinase(MAPK).lsm14 A,a key component of P-bodies and SGs,is still lacking in glioma.Therefore,a systematic analysis of the prognostic value of LSM14 A expression in glioma would be beneficial to gain insight into the molecular biological mechanisms of glioma and find therapeutic targets.Based on this,we analyzed the expression and prognostic differences of LSM14 A in glioma using the glioma public database,aiming to explore the role of LSM14 A in glioma development in order to find new therapeutic targets in glioma in the future.Methods: 1.To analyze the differential expression of LSM14 A in TCGA,CGGA,GEO and CPTAC databases using bioinformatics techniques and further analyze the correlation between LSM14 A and patient prognosis.2.Use qPCR and Western Blot to detect the expression level of LSM14 A in clinical glioma samples.3.To construct stable transgenic cell lines with overexpression and knockdown of LSM14 A,and to detect the m RNA and protein expression of LSM14 A by qPCR and Western Blot.4.In vitro CCK8,Ed U,clonal colony formation assay and Transwell migration assay were performed to detect the effect of LSM14 A on the proliferation and migration of glioma cells.5.Western Blot to detect the effects of LSM14 A knockdown and overexpression on the expression of cell cycle-related factors.Results: 1.Bioinformatics analysis of TCGA,CGGA,GEO,and CPTAC databases showed that LSM14 A m RNA and protein expression levels were higher in gliomas than in normal tissues,and increased in gliomas with higher malignancy.lsm14 A expression was elevated in IDH wild-type and 1p/19 unco-deficient gliomas,and high expression of LSM14 A was positively correlated with poor prognosis in glioma patients.patients had a positive correlation with poor prognosis.The difference was statistically significant,P <0.05.The expression levels of LSM14 A in clinical glioma samples by qPCR and Western Blot were higher in clinical GBM samples than in LGG samples,and the differences were statistically significant,P < 0.05.3.Stable transglioma cell lines with knockdown and overexpression of LSM14 A were constructed.qPCR and Western Blot verified that the knockdown efficiency was more than0.5 and overexpression efficiency was more than 1.5-fold.4.In vitro CCK8,Ed U,and clonal colony formation assays demonstrated that LSM14 A promoted the proliferation of glioma cells,P < 0.01,and Transwell assay demonstrated that LSM14 A promoted the migration of glioma,P < 0.001.5.Western Blot assay of LSM14 A knockdown and overexpression cell lines showed that protein expression of cell cycle inhibitory factor P21 was negatively correlated with LSM14 A,and protein expression of cell cycle promoting factor CDK4 was positively correlated with LSM14 A.Conclusion: In this study,we found that LSM14 A was highly expressed in glioma and increased with glioma grade;high expression of LSM14 A was positively correlated with poor prognosis of glioma patients;LSM14A promotes proliferation and migration of glioma cells and may affect the proliferation of glioma cells by regulating cell cycle-related molecules P21 and CDK4.