| Research Background:Multiple myeloma(MM)is a kind of malignant clonal plasma cell disease with high cytogenetic heterogeneity.As a malignant disease of cancer,it is mainly caused by the malignant proliferation of plasma cells in bone marrow,which belongs to the category of malignant diseases of blood system.The incidence of MM is the second highest in multiple myeloma,a malignant disease of the blood system.As a senile disease,the peak age of MM is 55-74 years old,often accompanied by a variety of complications.In China,with the aging of the population,the incidence of MM has been increasing year by year.Along with the advance of genetic testing technology,more and more evidence shows that multiple myeloma has the decisive factor for the disease prognosis and outcome of genetics,the current results for MM prognosis assessment in genetics stratification treatment,but the MM at present is still an incurable disease,most patients will eventually progress to relapse refractory period.As is known to all,the occurrence and development of cancer is a complex regulatory process involving a variety of genes or proteins.With the continuous development of molecular biotechnology,more and more studies on genes have shown that long noncoding RNA(lncRNAs)in non-coding RNA plays a particularly important role in the pathogenesis of malignant tumor diseases.Abnormal expression of lncRNAs may lead to various hematologic malignancies,including lymphoma,leukemia and MM.Studies on MM have shown that there are many different abnormal expressions of lncRNAs at different stages of MM development.These lncRNAs are closely related to the evolution of MM and have been proved to regulate the expression level of genes or proteins,thus being targeted for research and playing a role in the occurrence and development of multiple myeloma.This study mainly discussed the role of lncRNA LINC01003 in multiple myeloma disease and the downstream regulation mechanism affecting the disease.Methods:1.Real-time quantitative polymerase chain reaction(QRT-PCR)was used to detect the expression levels of lncRNA LINC01003,Mir-33A-5p and PIM1.2.Cell lines with different gene expression levels were constructed by cell culturerelated methods such as cell culture,passage,transfection and cryopreservation;3.Western blot assay(WB)was used to detect the relative protein expression levels of apoptosis related factors(Bcl-2 and Bax).4.The cell viability was detected by MTT colorimetry.5.The protein expression levels of adhesion related proteins(muc-1,VCAM-1 and ICAM-1)were detected by enzyme-linked immunosorbent assay(ELISA).6.The targeting relationship between LINC01003,Mir-33a-5p and PIM1 was detected by double luciferase reporting assay,RNA immunoprecipitation(RIP)assay and RNA pull-down assay,respectively.Results:1.The expression level of lncRNA LINC01003 in peripheral blood of patients with multiple myeloma is lower than that of normal healthy adults.2.In patients with multiple myeloma,the expression level of lncRNA LINC01003 was correlated with ISS stage,and the later stage,the lower the expression level.3.Both upregulation of lncRNA LINC01003 and knockdown of Mir-33a-5p inhibited the survival rate and adhesion of multiple myeloma cell lines,and promoted apoptosis.4.Mir-33a-5p was confirmed as the downstream target of lncRNA LINC01003,and PIM1 was confirmed as the direct target gene of Mir-33a-5p.5.Both the high expression of Mir-33a-5p and the low expression of PIM1 can reverse the inhibition of the adhesion and survival rate of multiple myeloma cell lines and the promotion of apoptosis caused by the overexpression of lncRNA LINC01003.Conclusion:LncRNA LINC01003 affects the expression of its downstream target gene Mir-33a-5p by virtue of its molecular "sponge" effect on Mir-33a-5p,thus achieving the inhibitory effect on multiple myeloma. |
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