Study On The Regulation Of Antisense RNA To The Transcription Of RB1Gene In NSCLC

Objective:To confirm that antisense RNA regulates the transcription of RB1genein NSCLC.Further more,that can provide reference material for further study on theregulation of antisense RNA to RB1gene in apparent genetic level.Moreover,our studywill provide experiment basis for the research of RB1gene and anti-RNA in humantumours.Methods:1.Extract RNA from four NSCLC cell strains containing H1299、SPC-A1、A549and H520,and clone RB1gene and its antisense transcript-ASRB1bythe method of Real Time fluorescence PCR.Finally, analyse the relative trancriptionlevels of the two.2.Extract RNA from the NSCLC cell line—SPC-A1,and carry out3’RACE.In theend, the3’ sequence of ASRB1can be known by sequencing and BLAST.3.Interfere ASRB1with siRNA in cell lines of H1299and H520,and detect themRNA and protein expression levels of RB1gene.Compare and analyse theexperimental group(interfered group) and control group(non-interfered group).Results:1.In the cell lines of H1299and H520，there’s a relatively higherexpression of RB1gene and lower expression of ASRB1.However,RB1gene presentsrelatively lower expression and ASRB1relatively higher expression in A549andSPC-A1cell lines.Moreover,there’s a negative correlation between the transcriptionallevel of RB1gene and ASRB1in one cell.（r2=0.891，P<0.01）.2.You can see the3’ sequence of ASRB1in the thesis.3.By RNA interference in H1299and H520cell lines,the mRNA and proteinexpression levels of RB1gene in the experimental group are higher than that in thecontrol group.Conclusion:1.There’s a different expression level among various NSCLC cell strains,but there’s a negative correlation between the transcriptional level of RB1geneand ASRB1in one cell.2.In order to design a siRNA specific to ASRB1,we succeed in getting the3’sequence of ASRB1.3.ASRB1has the negative regulation of the transcription of RB1gene in NSCLC.