RNA干扰LOXL2基因对头颈部鳞状细胞癌凋亡及PD-L1表达的影响

目的:探讨抑制赖氨酰氧化酶样蛋白2(LOXL2)基因表达对头颈部鳞状细胞癌(HNSCC)增殖凋亡及PD-L1表达的影响研究。方法:通过Western blot检测人HNSCC细胞YCU-H891、YCU-N861和KB中LOXL2的蛋白表达。以Lipo-fectamineTM2000为载体,参照其说明将合成的LOXL2 siRNA转染KB细胞,转染48 h后,Western blot检测LOXL2、PD-L1、STAT3、p-STAT3、PCNA和Bax的蛋白表达; CCK8及流式细胞仪分别检测细胞活力及凋亡率。结果:LOXL2在YCU-H891、YCU-N861和KB中表达均升高。转染LOXL2 siRNA的KB细胞LOXL2的蛋白表达明显降低,与空白组和NC组比较差异均具有统计学意义(P<0. 05)。转染LOXL2 siRNA的KB细胞活力明显降低,凋亡率明显升高,PD-L1、p-STAT3和PCNA的蛋白表达均明显降低,Bax的蛋白表达明显升高,与NC组比较,差异均具有统计学意义(P<0. 05)。结论:RNA干扰抑制LOXL2基因可降低HNSCC细胞活力和诱导凋亡,下调PD-L1表达,机制可能与抑制STAT3信号通路有关。

Effect of inhibition of LOXL2 gene by RNA interference in head and neck squamous cell carcinoma apoptosis and PD-L1 expression

Objective:To investigate the effect of LOXL2 gene expression was inhibited on proliferation,apoptosis and PD-L1 expression in the head and neck squamous cell carcinoma(HNSCC).Methods:LOXL2 protein expression in human HNSCC cells YCU-H891,YCU-N861 and KB were detected by Western blot.Lipofectamine TM 2000 was used as the carrier,the LOXL2 siRNA was transfected into KB cells according to its instructions.Cells were transfected for 48 h,the expression of LOXL2,PD-L1,STAT3,p-STAT3,PCNA protein were detected by Western blot,and the cell viability and apoptosis rate were detected by CCK8 and flow cytometry.Results:LOXL2 expression was increased in YCU-H891,YCU-N861 and KB cells.The protein expression of LOXL2 in KB cells transfected with LOXL2 siRNA decreased significantly compared with blank group and NC group(P<0.05).The activity of KB cells transfected with LOXL2 siRNA decreased obviously,the apoptosis rate increased obviously,the protein expression of PD-L1,p-STAT3 and PCNA decreased obviously,and the protein expression of Bax increased obviously,the difference was statistically significant compared with NC group(P<0.05).Conclusion:Inhibition of LOXL2 gene expression by RNA interference can reduce HNSCC cell viability and induce apoptosis,and down regulate PD-L1 expression.The mechanism may be related to inhibition of STAT3 signaling pathway.