Studies On The Expression And Significance Of ACSS2 In Non-Small Cell Lung Cancer

Objectives:This study intends to identify ACSS2 expression in non-small cell lung cancer and explore the effect on cellular proliferation,apoptosis and migration of A549 cells by RNA interference.It may provides new basis for the follow-up experiments of ACSS2 and offers a new therapy method for non-small cell lung cancer.Methods:The NSCLC tissues and corresponding adjacent normal tissues were collected and conserved.RT-PCR was used to employed ACSS2 expression in NSCLC tissues.The ACSS2 interference fragment ACSS2-siRNA and negative control was designed and synthesis for RNA interference.The transient transfection was proceed in non-small cell lung cancer cell line A549.Fluorescence based quantitative polymerase chain reaction was used to detection ACSS2 mRNA expression.MTT,flow cytometry and wound healing assay was used to detect cell proliferation,apoptosis rate and migration.Results:1.the expression of ACSS2 in Non-small cell lung cancer and normal lung tissuesFluorescence based quantitative polymerase chain reaction was used to detect ACSS2 mRNA in non-small cell lung cancer and normal lung tissue.There are 77%(23/30)has high expression of ACSS2 in lung cancer group compared with the normal lung tissue.This difference has statistical significance meaning(P<0.05).2.The ACSS2 mRNA expression in A549 cells after ACSS2-siRNA transfectionACSS2-siRNA and control siRNA(control group)were transfection to A549cells through LipofectamineT~M2000.QRT-PCR was used to detect ACSS2 mRNA changes in the cells after transfection.The results show that ACSS2 mRNA was significantly lower expression after transfection the interferencefragment ACSS2-siRNA in non-small cell lung cancer cell line A549.This difference has statistical significance meaning(P<0.05).3.The inhibition of cell growth after transfect ACSS2-siRNAMethyl thiazolyl tetrazolium(MTT)was applied to detect the growth inhibition of non-small cell lung cancer cell line A549 after transfection ACSS2-siRNA 24,48,72,96 hours.The results showed that the growth inhibition rate at 24,48,72,96 hours after transfection siACSS2 were(8.5�5.2)%,(17.3�3.6)%,(22.9�4.1)%and(29.7�6.5)%respectively.Compared with the control group,there were no difference obvious(P=0.06)after 24 hours.Obvious difference with statistical significance(P<0.05)was observed after 48,72,96 hours.The Knock down of ACSS2 can obviously inhibit cell proliferation of non-small cell lung cancer cell line A549.4.The effects on apoptosis of A549 cells after transfect ACSS2-siRNAFlow cytometry was apllied to test influence on apoptosis of A549 cells between ACSS2-siRNA transfection and untreated groups.ACSS2-siRNA transfection group cell apoptosis rate was increased obviously compared with negative control and untreated group.Early apoptosis rate of three groups were(21.6�2.92)%,(5.7�2.51)%,(8.5�1.60)%respectively.Early apoptosis rate of ACSS2-siRNA transfection group was obviously higher than that of the negative and blank control group(P<0.05),and no obvious difference between the rear two groups.The results show that the knock down of ACSS2 of A549 cells can promote the early apoptosis.5.The influence of invasion force of A549 cells after transfect ACSS2-siRNAWound healing assay was used to detect cell migration ability.The results were analysed by Image-Pro plus 6.0.Measuring the migration distance of A549 cells after transfect ACSS2-siRNA 0,24,48hours.The cell migration distance of ACSS2-siRNA transfection group,negative control group and blank control group in24h were about(32.6�5.8)um,(44.6�8.4)um and(42.9+8.0)um respectively.There was no statistically significant difference among three groups(P>0.05,P=0.87).At 48 h the cell migration was(83.1�7.7)um,negative control group(120.3�8.3)um,the blank control group(125.5�7.6)um.Compared with the three groups,there have obvious statistically significant difference in cell migration distance(P<0.05).Conclusion:Knockdown the ACSS2 mRNA of non-small cell lung cancer cell line A549 can significantly inhibit the cell proliferation,migration ability and promote the early phase apoptosis rate.This study indicate that ACSS2 may contribute to the progression of human NSCLC.