实时定量PCR检测镉对人肝癌细胞SMMC-7721中TRPC1和TRPC4 mRNA表达的影响

目的：观察氯化镉（CdCl₂）处理后人肝癌SMMC-7721细胞中TRPC1和TRPC4 mRNA表达的改变，在mRNA水平探讨镉对肝癌细胞钙通道的影响。方法：以人肝癌细胞株SMMC-7721为研究模型，细胞暴露于氯化镉终浓度为0、5、10、20、30及40 μmol·L^-1的培养液中24 h。采用SYBR荧光实时定量PCR法检测TRPC1和TRPC4 mRNA表达，相对定量采用比较CT值法。结果：随着剂量的增加，TRPC1 mRNA表达有降低的趋势，除20 μmol·L^-1组外，各组mRNA表达均高于对照组（P<0.01）；TRPC4 mRNA表达有先增高后降低的趋势，5、10和30 μmol·L^-1组mRNA表达均高于对照组（P<0.01）。2个基因均在低剂量（5和10 μmol·L^-1）组表达增加幅度较大（P<0.01）。结论：低剂量镉可显著增加人肝癌SMMC-7721细胞中TRPC1和TRPC4的表达，镉可在mRNA水平对SMMC-7721细胞钙通道产生影响。

Effect of cadmium on expressions of TRPC1 and TRPC4 mRNA in SMMC-7721 cells detected with quantitative PCR

Objective To study the effects of cadmium chloride(CdCl2) on calcium channel by observing the expressions of TRPC1 and TRPC4 mRNA in SMMC-7721 cells.Methods SMMC-7721 cell cultures were exposed to cadmium at concentrations of 0,5,10,20,30 and 40 μmol·L-1 for 24 h.Power SYBR Green PCR Master Mix and Mx3000P QPCR System were used to measure a threshold cycle(CT) value for each sample.Relative quantitation of gene expression was based on the comparative CT method.Results The TRPC1 mRNA level tended to decrease along with the elevating of cadmium dose.The expressions of TRPC1 mRNA in all experimental groups were significantly higher than that in control group(P<0.01),except 20 μmol·L-1 group.The TRPC4 mRNA level first increased then decreased along with the elevating of cadmium dose.The expressions of TRPC4 mRNA in 5,10 and 30 μmol·L-1 groups were significantly higher than that in control group(P<0.01).Conclusion Cadmium can affect the calcium channels by inducing the expressions of TRPC1 and TRPC4 mRNA in SMMC-7721 cells at low doses.