CAJ | 학술논문

目的探讨益气解毒方对CNE2Z鼻咽癌细胞迁徙运动潜能的影响及其与细胞骨架蛋白系统表达活性的关系.方法以正常大鼠血浆为空白对照,以益气解毒方大鼠血浆为干预因素,采用划痕法评价含药血浆对靶细胞CNE2Z鼻咽癌细胞迁徙运动潜能的影响,以扫描电镜观察干预前后靶细胞伪足形态特征,免疫组化SABC法检测β-tubulin、F-actin、p-Ezrin、Rac1表达活性,RT-PCR法检测p-Ezrin mRNA转录活性,评价靶细胞迁徙运动潜能变化与细胞骨架蛋白系统表达活性的关系及其对益气解毒方干预作用的反应性.结果益气解毒方对CNE2Z鼻咽细胞的迁徙运动潜能显示抑制效应,在IC50浓度时,迁徙距离为(25.80 ±8.07) μm,抑制率为46.59％；靶细胞表现伪足变短小,伸展长度变短；β-tubulin、F-actin、p-Ezrin、Rac-1蛋白表达受抑制,图像灰度值分别为(0.857±0.059,0.900±0.040,0.812±0.050,0.785±0.022),与对照组的差异有统计学意义(P＜0.01)；p-Ezrin mRNA表达水平呈现同样变化趋势.结论益气解毒方对CNE2Z细胞的迁徙运动潜能发挥抑制效应,伴随以靶细胞骨架蛋白系统生物活性的降低,细胞伪足伸展抑制.
목적 탐토익기해독방대CNE2Z비인암세포천사운동잠능적영향급기여세포골가단백계통표체활성적관계.방법 이정상대서혈장위공백대조,이익기해독방대서혈장위간예인소,채용화흔법평개함약혈장대파세포CNE2Z비인암세포천사운동잠능적영향,이소묘전경관찰간예전후파세포위족형태특정,면역조화SABC법검측β-tubulin、F-actin、p-Ezrin、Rac1표체활성,RT-PCR법검측p-Ezrin mRNA전록활성,평개파세포천사운동잠능변화여세포골가단백계통표체활성적관계급기대익기해독방간예작용적반응성.결과 익기해독방대CNE2Z비인세포적천사운동잠능현시억제효응,재IC50농도시,천사거리위(25.80 ±8.07) μm,억제솔위46.59％；파세포표현위족변단소,신전장도변단；β-tubulin、F-actin、p-Ezrin、Rac-1단백표체수억제,도상회도치분별위(0.857±0.059,0.900±0.040,0.812±0.050,0.785±0.022),여대조조적차이유통계학의의(P＜0.01)；p-Ezrin mRNA표체수평정현동양변화추세.결론 익기해독방대CNE2Z세포적천사운동잠능발휘억제효응,반수이파세포골가단백계통생물활성적강저,세포위족신전억제.
Objective To investigate the intervening effect of Qi-Boosting Toxin-Resolving Formula (QBTRF) on the migration potentiality of nasopharyngeal carcinoma (NPC) cells and its association with the expressive activities of cellular skeletal protein system based on an experimental study in vitro.Methods In this study,QBTRF medicine-containing plasma of rats was taken as the intervening factor to treat the targeted NPC cells of CEN2Z for evaluating the correlation of changes in migrating potentiality of CEN2Z cells with expression activities of cellular skeletal protein system and their response to the pharmacological intervening effects of QBTRF,compared with the normal plasma of rats as the control.Here,scratching test was taken to evaluate the migration potentiality of targeting cells,immunohistochemical SABC assays were used to determine the expression activities of β-tubulin,F-actin,p-Ezrin and Rac1,which were taken as the typical indicators of cellular skeletal protein system.Scanning electron microscopy was selected to observe the changes in cellular pseudopodia.Reverse transcription-polymerase chain reaction (RT-PCR) procedure was used to detect the transcriptional activity of p-Ezrin mRNA.Results QBTRF showed a very significant inhibitory effect on the migration potentiality of CEN2Z cells,with its migration distance reduced to (25.80 ±8.07) μm and an inhibitory rate reached at 46.59％ at the concentration of IC50.Meanwhile,their cellular pseudopodia was shortened significantly,accompanied with very reduced extension length.Obviously decreased were the expression activities of β-tubulin,F-actin,p-Ezrin and Rac1 proteins,with their figure gray values being 0.857 ±0.059,0.900 ±0.040,0.812 ±0.050 and 0.785 ±0.022,respectively,with statistical significance compared with controls (P ＜0.01).Also seen was the same tendency of transcriptional level of p-Ezrin mRNA when cells cultured with QBTRE medicines-containing plasma together.Conclusions QBTRF holds significantly inhibitory effect on the migration potentiality of CEN2Z NPC cells.This kind of inhibitory effect may be associated with the reduced biological activity of cellular skeletal protein system and corresponding cellular pseudopodia changes.