Expression of long chain non coding RNA tumor protein 53 target gene 1 in skin basal cancer cells and its relationship with proliferation
Objective To investigate the expression of long non-coding RNA TP53 target gene 1(lncRNA TP53TG1)in human skin basal carcinoma cells and its relationship with proliferation.Methods The expression levels of lncRNA TP53TG1 were analyzed using real-time fluorescence quantitative polymer-ase chain reaction(PCR)in normal human skin cells(HaCaT)and human basal cell carcinoma TE 354.T and A431 cell lines.The highly expressed skin basal cell carcinoma TE 354.T cells were selected as the research objects,and infected with lentivirus to establish lncRNA TP53TG1 overexpressing cell lines and control cells,named as the control group and TP53TG1 group,respectively.The proliferation ability of cells in the control group and TP53TG1 group was analyzed using cell counting kit-8(CCK-8)and clone formation experiments.The expression levels of proliferation cell nuclear antigen(Ki-67)and proliferating cell nuclear antigen(PCNA)in two groups were analyzed by Western blotting.The proliferation activity of two groups was measured using the 7-aminoactinomycin D(7-AAD)assay kit.The in vitro tumor transplan-tation model was prepared in nude mice.The tumor volume and weight of control group and TP53TG1 group were analyzed.The gene levels of lncRNA TP53TG1 downstream related to cell proliferation were analyzed by real time fluorescence quantitative PCR.The comparison of inter group measurement data was done by t-test.Results The expression level of TP53TG1 in normal human skin cells(HaCaT)(0.83±0.34)was significantly lower than that in TE 354.T and A431 cell lines(1.65±0.20,1.58±0.13,t=5.579,5.503,P＜0.05).The 24-h absorbance value in the TP53TG1 group(2.06±0.12)was significantly higher than that in the control group(1.50±0.08,t=10.070,P＜0.05).The 14-day clone formation rate in the TP53TG1 group[(81.86±8.71)％]was significantly higher than that in the control group[(54.85±6.79)％,t=6.469,P＜0.05].The positive rate of 7-AAD in the TP53TG1 group[(65.57± 5.62)％]was significantly higher than that in the control group[(37.43±4.50)％,t=10.340,P＜0.05].The expression levels of Ki-67 and PCNA proteins in the TP53TG1 group(1.31±0.14,1.38± 0.15)were significantly higher than those in the control group(0.70±0.08,0.72±0.08,t=10.240,10.150,P＜0.05).The volume and mass of tumors formed subcutaneously in the TP53TG1 group[(986.71±81.64)mm3,(3.45±0.36)g]were significantly increased as compared with those in the control group[(699.29±53.62)mm3,(2.15±0.26)g,t=7.786,7.709,P＜0.05].The levels of microRNA(miR)-33a-5p and miR-33b in the TP53TG1 group(0.72±0.10,0.65±0.06)were signifi-cantly higher than those in the control group(0.99±0.30,1.03±0.12,t=2.227,7.589,P＜0.05).Conclusion The expression of lncRNA TP53TG1 in human skin basal cancer cells significantly de-creased.Overexpression of lncRNA TP53TG1 can significantly inhibit the proliferation of human skin stro-mal cancer cells,which may be related with miR-33a-5p and miR-33b.
Long chain non coding RNATumor protein 53 target gene 1Skin basal cancer cellsProliferation
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