Investigation Of The Effect And Its Mechanism Of LncRNA03882 On The Migration And Invasion Of Ovarian Cancer Cells

Objective:To investigate the effect and of Long non-coding RNAs,differentially expressed in ovarian cancer,on migration and invasion of ovarian cancer cells and its molecular mechanism via bioinformatics analysis and cellular and molecular biological methods.Methods:（1）CRC（Coding Potential Caculator:http://cpc.cbi.pku.edu.cn/）,an online worker of bioinformatics,was used to analyze the non-coding ability of Lnc RNA03882.（2）Bioinformatics online tool Annolnc（http://annolnc.cbi.pku.edu.cn/）was performed to analyze the m RNA expression of Lnc RNA03882 in tumor tissues and normal tissues.（3）The genes that may relate to the m RNA expression level of Lnc RNA03882 were screened by Person correlation analyze,and the intensity of co-expression was expressed by Pearson correlation coefficient.（4）The genes with high Pearson correlation coefficient were selected and analyzed by bioinformatics for gene ontology analysis（gene ontology,GO analysis）and related signal pathway analysis（Kyoto Encyclopedia of Genes and Genomes,KEGG）.（5）The expression of Lnc RNA03882 in ovarian cancer cells was confirmed by q RT-PCR.（6）Specific si RNA was used to knock down the expression of Lnc RNA03882 in SKOV3 cells.（7）The effect of Lnc RNA03882 on the migration and invasion of SKOV3 cells was detected by transwell migration and invasion assay.（8）The location of Lnc RNA03882 was analyzed by Lnc Locater database.（9）Fluorescence in situ hybridization（FISH）was used to verify the localization of Lnc RNA03882.（10）q RT-PCR and Western Blot were used to detect the m RNA and protein expression of zinc finger E-box binding protein 1（Zinc Finger E-Box Binding Homeobox 1,ZEB1）after knocking down Lnc RNA03882.（11）RIP technique was performed to illustrate the interaction between Lnc RNA03882and ZEB1.（12）Western Blot was used to detect the expression of E-cadherin(E-Ca²⁺dependent cell adhesion molecules family,E-cadherin)in SKOV3 cells after knocking down Lnc RNA03882.（13）Based on the database of TCGA,Kaplan-Meier survival analysis was used to study the prognostic effect of Lnc RNA03882 on ovarian cancer.Result:（1）In bioinformatics analysis,the score for predicting the coding ability of Lnc RNA03882 in CRC was-1.12117,and when the value of Lnc RNA03882 is less than 0,there is no coding conservative.（2）The expression of FPKM（Fragments Per Kilobase Million,FPKM）of Lnc RNA03882 in ovarian cancer tissues was higher than that in normal ovarian tissues.（3）Among the genes related to Lnc RNA03882,there were 207 co-expressed genes which Pearson correlation coefficient≥0.8.（4）GO analysis of 207 related genes and Lnc RNA03882 was mainly enriched in the process of cell cycle regulation,cell migration and invasion,and KEGG pathway analysis was mainly concentrated in cell cycle,mitosis and so on.（5）q RT-PCR detection showed that Lnc RNA03882 was highly expressed in ovarian cancer cell lines and low in normal tissues.（6）Specific si RNA decreased the expression of Lnc RNA03882 in SKOV3 cells.（7）When Lnc RNA03882 was knocked down,the migration and invasion ability of SKOV3 decreased.（8）Bioinformatics analysis of Lnc Locater database predicted and declared that Lnc RNA03882 was located in the cytoplasm.（9）FISH assay showed that Lnc RNA03882 was expressed in both cytoplasm and nucleus,but the expression level in cytoplasm was higher than that in nucleus.（10）After knocking down Lnc RNA03882,both m RNA and protein expression levels of ZEB1 were down-regulated.（11）RIP showed that Lnc RNA03882 can be directly combined with ZEB1.（12）ZEB1 encodes a transcriptional inhibitor of E-cadherin.After knocking down Lnc RNA03882,the level of ZEB1 protein was down-regulated and the expression of E-cadherin was up-regulated.（13）The survival rate of patients with high expression of Lnc RNA03882 was significantly lower than that of patients with high expression of Lnc RNA03882.Conclusion:Based on investigation of bioinformatics,cellular and molecular biological methods,it is found that the expression of Lnc RNA03882 is upregulated in ovarian cancer.It participates in a variety of biological processes of ovarian cancer,which is related to the migration,invasion and prognosis of ovarian cancer.It may become a new biomarker of ovarian cancer in the future.