摘要
【目的】探讨p75神经营养因子受体(p75 neurotrophin receptor,p75NTR)对口腔鳞状细胞癌细胞迁移、侵袭能力的影响及机制。【方法】以口腔鳞状细胞癌细胞Tca8113为研究对象,流式细胞术筛选p75NTR阳性细胞,细胞转染小干扰RNA阴性对照(siRNA control)和p75NTR小干扰RNA(p75NTR siRNA)分别命名为NC组和干扰组,同时以没有转染的细胞为未转染组,RT-PCR和Western blotting检测转染后细胞中p75NTR mRNA和蛋白水平,细胞划痕实验检测细胞迁移,Transwell小室检测细胞侵袭,Western blotting检测细胞中基质金属蛋白酶-2(matrix metalloprotease,MMP-2)、基质金属蛋白酶-9(matrix metalloprotease 9,MMP-9)、蛋白激酶B(protein kinase B,Akt)、磷酸化蛋白激酶B(p-Akt)蛋白水平。【结果】未转染组、NC组、干扰组p75NTR mRNA表达水平依次为:(1.02±0.09)、(0.99±0.05)、(0.36±0.06),p75NTR蛋白水平依次为:(0.86±0.06)、(0.86±0.07)、(0.32±0.04)。干扰组p75NTR mRNA和蛋白水平均明显低于未转染组(P<0.05)。未转染组、NC组、干扰组细胞迁移率依次为:(100.01±11.36)%、(100.03±10.94)%、(62.94±5.74)%。干扰组细胞迁移率明显低于未转染组(P<0.05)。未转染组、NC组、干扰组侵袭细胞数目依次为:(115.64±13.05)、(114.87±14.61)、(73.25±9.33)。干扰组侵袭细胞数目明显低于未转染组(P<0.05)。未转染组、NC组、干扰组细胞中MMP-2蛋白水平依次为:(1.25±0.09)、(1.26±0.11)、(0.42±0.03),MMP-9蛋白水平依次为:(0.53±0.04)、(0.52±0.07)、(0.20±0.04),Akt蛋白水平依次为:(1.05±0.06)、(1.02±0.08)、(1.05±0.04),p-Akt蛋白水平依次为:(0.28±0.04)、(0.27±0.06)、(0.02±0.01)。干扰组细胞中MMP-2、MMP-9、p-Akt表达水平均明显低于未转染组(P<0.05)。【结论】干扰p75NTR表达后的口腔鳞状细胞癌细胞侵袭迁移能力下降,其作用机制可能与细胞内MMP-2、MMP-9和Akt信号通路有关。
[Objective]To explore the effects of p75 neurotrophin receptor （p75NTR） on the migration and invasion in oral squamous cell carcinoma and the related mechanism. [Methods] The cell line Tca8113 of oral squamous cell carcinoma was used as the subject in this study. The p75NTR positive cells were screened by flow cytometry. The cells were transfected with siRNA control in the NC group and p75NTR siRNA in the interference group, respectively, while the cells in the non transfection group were not transfected. The mRNA and protein levels of p75NTR in transfected cells were detected by RT-PCR and Western blotting. Cell scratch test was used to detect cell migration, and cell invasion was detected by Transwell chamber. The protein levels of matrix metalloprotease （MMP）- 2, MMP-9, protein kinase B （Akt） and p-Akt were detected by Western blotting. [ Results] The mRNA expression levels of p75NTR in the non transfection group, NC group and interference group were （1.02 ± 0.09）, （0.99 ± 0.05） and （0.36 ± 0.06） respectively, and the protein levels of p75NTR were （0.86 ± 0.06）, （0.86 ± 0.07） and （0.32± 0.04） respectively. The mRNA and protein levels of p75NTR in the interference group were significantly lower than those in the non transfection group （P〈O.05）. The rates of cell migration in the non transfection group, NC group and interference group were （100.01 ± 11.36）%, （100.03 ± 10.94）% and （62.94 ± 5.74）%. The rate of cell migration in the interference group was obviously lower than that in the non transfection group （P〈0.05）. The number of invasive cells in the non transfection group, NC group and interference group was （115.64 ± 13.05）, （114.87 ± 14.61） and （73.25 ± 9.33） respectively. The number of invasive cells in the interference group was significantly lower than that in the non transfection group （P〈0.05）. The protein levels of MMP-2 in the non transfection group, NC group and interference group were （1.25 ± 0.09）, （1.26 ± 0.11） and （0.42 ± 0.03） respectively; the protein levels of MMP-9 were （0.53 ± 0.04）, （0.52 ± 0.07）, （0.20 ± 0.04）respectively; the protein levels of Akt were （1.05 ± 0.06）, （1.02 ± 0.08） and （1.05 ± 0.04） respectively; the protein levels of p-Akt were （0.28 ± 0.04）, （0.27 ± 0.06） and （0.02 ± 0.01） respectively. The expression levels of MMP-2, MMP-9 and p-Akt in the interference group were significantly lower than those in the non transfection group （P〈0.05）. [Conclusion ] The invasion and migration ability of oral squamous cell carcinoma ceils decreases after the interference in the expression of p75NTR, and its mechanism may be related to the intracellular MMP-2, MMP-9 and Akt signaling pathway.
出处
《武警后勤学院学报（医学版）》
CAS
2017年第8期657-661,共5页
Journal of Logistics University of PAP（Medical Sciences）
基金
海南省自然科学基金项目(814315)
