5-氨基酮戊酸-光动力治疗宫颈癌体外实验分析

Analysis of 5-amino-ketovalerate in vitro with photodynamic treatment of cervical cancer

目的分析对宫颈癌Hela细胞采用5-氨基酮戊酸-光动力治疗对其Toll样受体4/核因子-kB(TLR4/NF-kB)信号通路的影响。方法将宫颈癌Hela细胞分为空白组、阴性siRNA组以及TLR4-siRNAs组,分别观察在562 nm红光照射20 min后其24 h、48 h、72 h时目标组与阴性组宫颈癌Hela细胞增殖、凋亡以及迁移改变情况。结果1.0 mmol/L浓度的ALA组低倍镜视野下细胞计数、细胞增殖率与空白组比较,差异有统计学意义(P<0.05)。1.0 mmol/L浓度的ALA组p65、p-p65、Bcl-2及TLR4-mRNA与空白组比较,差异有统计学意义(P<0.05)。实验目标组TLR4-siRNAs组采用ALA最适浓度对宫颈癌Hela细胞与空白组siRNA阴性p65、Bcl-2的mRNA水平为(1.03±0.16)mmol/L、(2.78±0.35)mmol/L,蛋白水平分别为(3.29±0.08)mmol/L、(4.10±0.23)mmol/L,对比差异有统计学意义(P<0.05)。TLR4-siRNAs组低倍镜视野下细胞计数、细胞增殖率与空白组、阴性siRNA组比较,差异有统计学意义(P<0.05)。结论通过对宫颈癌细胞采用5-氨基酮戊酸-光动力疗法,有效的抑制其TLR4/NF-κB信号通路,抑制肿瘤细胞的转录与增殖同时增加肿瘤细胞的凋亡能力。

Objective To analyze the effect of 5-amino-ketovalerate photodynamic therapy on toll-like receptor 4/nuclear factor-kB(TLR4/NF-kB)signaling pathway in Hela cells of cervical cancer.Methods Cervical cancer Hela cells transfected in vitro were divided into blank group,negative siRNA group and TLR4-siRNAs group.The proliferation,apoptosis and migration of cervical cancer Hela cells in target group and negative group were observed at 24 h,48 h and 72 h after 20 min of red light irradiation at 562 nm,respectively.Results The cell count and cell proliferation rate in the 1.0 mmol/L concentration ALA group were significantly different from those in the blank group at low magnification(P<0.05).There were significant differences in p65,P-P65,Bcl-2 and TLR4-mRNA in ALA group with 1.0 mmol/L concentration compared with that in blank group(P<0.05).The mRNA levels of siRNAs negative p65 and Bcl-2 in the target group TLR4-siRNAS group were(1.03±0.16)mmol/L and(2.78±0.35)mmol/L in Hela cells compared with those in the blank group when the optimal concentration of ALA was applied.The protein levels were(3.29±0.08)mmol/L and(4.10±0.23)mmol/L,respectively,and the difference was statistically significant(P<0.05).The cell count and cell proliferation rate in TLR4-siRNAs group were statistically significant compared with blank group and negative siRNA group at low magnification field(P<0.05).Conclusion By using 5-amino-ketovalerate photodynamic therapy on cervical cancer cells,TLR4/NF-kB signaling pathway can be effectively organized,inhibit the transcription and proliferation of tumor cells,and increase the apoptosis capacity of tumor cells.