| Background:Multiple myeloma(MM)is a malignant proliferative disease that is common in middle-aged and elderly people and has abnormal plasma cells in the bone marrow.With the aging of China’s population and the development of myeloma diagnosis technology,the incidence of the disease in China has shown an upward trend,and the incidence has a tendency to become younger.Despite the emergence of a large number of new clinical first-line treatment drugs,the survival time of patients has been significantly improved,but due to drug resistance and relapse problems,multiple myeloma is still in an incurable state.Therefore,to study the formation and evolution of myeloma from the molecular level and the genetic level,to help provide the key theoretical basis and practical value in the prevention and treatment of the disease.Long non-coding RNA(long non-coding RNA,RNA)belongs to large-molecule non-coding RNA.It refers to a class of RNA molecules that are longer than 200 bases and have multiple regulatory functions.It is transcribed from RNA polymerase ∥.After cutting and processing,mature lncRNA is formed,but it will not be translated into protein.LncRNA regulates genes in terms of epigenetic regulation,transcription level and even post-transcription level.MicroRNA(microRNA,miRNA)is a type of non-coding single-stranded small RNA that is widely expressed in prokaryotes and eukaryotes and is approximately 20 bases in length.In recent years,studies have found that competitive endogenous RNA(ceRNA)can be used as a potential therapeutic target and biomarker when studying the pathogenesis of malignant tumors,which shows the extremely important clinical application value and Significance.At the same time,there is also a large amount of evidence that the circulating serum or plasma lncRNA can be used as a potential therapeutic target for malignant tumors,helping the diagnosis of malignant tumors,prompting prognosis and other conditions,and predicting the individual’s response to drug treatment and drug resistance problems or even malignant tumor Theexploration of formation mechanism pointed out the direction.purpose:Identify the characteristic long noncoding RNA(RNA)in multiple myeloma,and detect the expression level of related lncRNA in myeloma,study its correlation with the clinicopathological characteristics of myeloma,and finally draw the right Thoughts and theoretical basis related to myeloma gene diagnosis and targeted therapy.Method:1.Download the initial and relapsed myeloma sample data in the relevant databases(TargetScan database and mirDB database),detect the corresponding differentially expressed miRNA(DE-miR),and then extract the lncRNA of interest in the relevant database(Differentially expressed long noncoding RNA,DE-lncRNA)and mRNA(differentially expressed messenger RNA,DEG),extract the lncRNA and mRNA of interest and the corresponding FPKM value,replace the 0 value with1/2 of the minimum value,and use log10(FPKM)Heat map with values,parameter scale = row.Then use TargetScan and mirDB to predict the miRNA target gene provided in the heat map,mirDB filter parameters(TargetScore> 80),and then take the intersection with the statistically significant difference mRNA in this sequencing result and make a venn map.Take the gene in the intersection of venn diagrams as the target gene,extract its corresponding miRNA(select the miRNA-mRNA interaction pairs predicted by both TargetScan and mirDB as the result of ceRNA analysis),and combine the selected lncRNA and its corresponding miRNA to select both control The miRNA whose mRNA controls lncRNA serves as the miRNA for the final ceRNA analysis.These co-corresponding expressed RNAs are studied in multiple databases to explore the relationship between them.Then,the mRNA-miRNA network and the lncRNA-miRNA network are constructed,and finally the two The authors integrated into the ceRNA network control map of lncRNA-miRNA-mRNA.Then we use Cytoscape software to visualize the ceRNA network control map,and use some important features(edge ? ? aggregation coefficient,Pearson correlation coefficient)that describe the network topology to filter out the relationship pairs in the ceRNA network map.2.Collect 20 cases of bone marrow of patients with multiple myelomadiagnosed in the First Affiliated Hospital of Nanchang University from January 2012 to October 2018 and 20 cases of bone marrow of the control group,extract 5 ml of bone marrow,and collect them with EDTA anticoagulation test blood collection tube.Total RNA was extracted and then reverse transcribed into cDNA.Real-time fluorescence quantitative PCR(qRT-PCR)technology was used to detect the expression of IRAIN in bone marrow and peripheral blood.At the same time,the correlation between the expression level of IRAIN and the corresponding clinical pathological characteristics was analyzed.The results were analyzed using statistical software SPSS 19.0.Result:1.We finally obtained 279 intersecting genes from the venn diagram,and selected 12 miRNAs that control both mRNA and lncRNA as the final miRNA analysis ceRNA analysis,they correspond to a total of 18 lncRNA and 118 mRNA2.The expression of LncRNA IRAIN detected in the plasma of myeloma patients is down-regulated compared with the normal control group(P <0.05).The expression level of IRAIN in the plasma of myeloma patients and the age,gender,international stage and classification of the corresponding patients There is no significant correlation between the type(P> 0.05),the expression level of IRAIN in the plasma of myeloma patients is closely related to the cytogenetic abnormalities of their corresponding patients(P <0.05),and the patients with cytogenetic abnormal-lities are more than those without.The expression level of LncRNA IRAIN is significantly reduced in patients Inconclusion:1.LINC01800 and LINC02357 are significantly overexpressed in the bone marrow tissue of the patient,but LIN104472848(IRAIN)is highly expressed in normal samples and significantly lower in the bone marrow tissue of the patient.Due to the abnormal expression of LNCRNA IRAIN,it is possible that this indicator is Pathogenesis and formation and development are closely related,and play a very significant role in regulating the molecular mechanism of myeloma recurrence.2.The expression level of LNCRNA IRAIN in myeloma plasma is significantly lower than that of the normal control group,indicating that IRAIN may be used as atumor suppressor gene to regulate the occurrence and development of myeloma,and the expression level of IRAIN in the body It is closely related to the presence of cytogenetic abnormalities in myeloma patients.The expression level of IRAIN in patients with myeloma with cytogenetic abnormalities is significantly reduced compared to the expression level without abnormalities,indicating that IRAIN may be involved in the tumor as an inhibitor.Formation process,and IRAIN may become a predictive biomarker for multiple myeloma and provide a therapeutic target for individualized treatment. |
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