The Effection Of RNASEH1-AS1 On The Proliferation And Migration Of HCT116 Cells

Objective:Based on the previous research results,this project selected the long non-coding RNA RNASEH1-AS1 as the research object,which was obviously highly expressed in colorectal cancer tissues and cells.This project will explore the effection of RNASEH1-AS1 on the proliferation and migration ability of colorectal cancer cells,then through RNA-seq and bioinformatics analysis,to explore the possible biological processes and signaling pathways involved in colorectal cancer.Methods:(1)Separate the Nuclear and cytoplasmic RNA of colorectal cancer HCT116 cells,and MALAT1 and ACTIN were used as reference for nuclear and cytoplasmic localization,respectively.The nucleoplasmic localization of RNASEH1-AS1 was detected by reverse transcription and real-time fluorescence quantitative PCR.(2)SiRNA specifically targeting RNASEH1-AS1 was designed and synthesized,and cell proliferation experiments were carried out on HCT116 cells transfected with siRNA.(3)HCT116 cells were transfected with siRNA and cell cycle analysis was performed by flow cytometry.(4)The effection of RNASEH1-AS1 on the migration ability of HCT116 cells was investigated through wound healing,and the cell healing rate was quantitatively analyzed.(5)After transfecting HCT116 cells with two siRNAs targeting RNASEH1-AS1 with different sequences,the RNA was extracted for RNA-seq,and analysis the results.Then,Use gene enrichment(GO)analysis and KEGG pathway analysis on the genes co-regulated by the two siRNAs.Results:(1)RNASEH1-AS1 is mainly localized in the cytoplasm.(2)When RNASEH1-AS1 be knockdowned,the proliferation and migration ability of HCT116 cells were reduced.(3)There were 181 genes were upregulated and 253 genes were downregulated.(4)Up-regulated genes were related to the humoral immune response,cell differentiation and B cell proliferation.Down-regulated genes are mainly related to metabolic pathways such as oxygen transport,redox reaction,and also related to the cancer,such as neural development,immune response,and cell proliferation.Conclusion:(1)Knocking down RNASEH1-AS1 reduced the proliferation and migration ability of colorectal cancer cells.(2)The gene group regulated by RNASEH1-AS1 is involved in the functions and signal pathways related to tumorigenesis.