CAJ | 학술논문

　　目的观察益气解毒法不同组方对CNE2Z鼻咽癌细胞迁徙运动潜能的影响并探讨其效应与相关基因表达活性变化的关系。方法依据益气解毒法组成原方并在此基础上依法构建改方，分别制备不同浓度含药血浆并作用于相同靶细胞CNE2Z鼻咽癌细胞，划痕法检测药物干预前后靶细胞的迁徙运动潜能，免疫组化法和RT-PCR法分别检测干预前后靶细胞nm23和c-myc蛋白表达水平和mRNA转录活性，比较分析组间差异。结果细胞迁徙运动能力检测结果显示，原方呈现抑制效应而改方表现则促进效应(P＜0.05)。免疫组化检测结果，在c-myc与nm23-H1蛋白表达活性上，原方对前者表达活性无明显影响但后者显著增强，差异均有统计学意义(P＜0.05)；改方则对c-myc蛋白表达显示促进效应，对nm23-H1蛋白表达显示抑制效应(P＜0.05)。RT-PCR检测结果则显示，原方及改方的高中剂量处理组细胞nm23-H1 mRNA表达显著升高(P＜0.05)，低剂量组则无明显变化(P＞0.05)；改方高剂量组c-myc mRNA表达水平明显高于对照组(P＜0.05)，原方组则未显示明显药理效应(P＞0.05)。结论益气解毒原方对CNE2Z细胞的迁徙运动潜能发挥抑制效应，而由剂量变化构建的改方虽然显示有促进效应，可能与其影响nm23-H1及c-myc表达活性及交互作用有关，还需要进一步研究证实。研究结果也对中药组方中剂量把握的重要性提供了初步的实验依据。
　　목적관찰익기해독법불동조방대CNE2Z비인암세포천사운동잠능적영향병탐토기효응여상관기인표체활성변화적관계。방법의거익기해독법조성원방병재차기출상의법구건개방，분별제비불동농도함약혈장병작용우상동파세포CNE2Z비인암세포，화흔법검측약물간예전후파세포적천사운동잠능，면역조화법화RT-PCR법분별검측간예전후파세포nm23화c-myc단백표체수평화mRNA전록활성，비교분석조간차이。결과세포천사운동능력검측결과현시，원방정현억제효응이개방표현칙촉진효응(P＜0.05)。면역조화검측결과，재c-myc여nm23-H1단백표체활성상，원방대전자표체활성무명현영향단후자현저증강，차이균유통계학의의(P＜0.05)；개방칙대c-myc단백표체현시촉진효응，대nm23-H1단백표체현시억제효응(P＜0.05)。RT-PCR검측결과칙현시，원방급개방적고중제량처리조세포nm23-H1 mRNA표체현저승고(P＜0.05)，저제량조칙무명현변화(P＞0.05)；개방고제량조c-myc mRNA표체수평명현고우대조조(P＜0.05)，원방조칙미현시명현약리효응(P＞0.05)。결론익기해독원방대CNE2Z세포적천사운동잠능발휘억제효응，이유제량변화구건적개방수연현시유촉진효응，가능여기영향nm23-H1급c-myc표체활성급교호작용유관，환수요진일보연구증실。연구결과야대중약조방중제량파악적중요성제공료초보적실험의거。
Objective To investigate the effect of different formulae composed from the therapeutic principle of Qi-boosting toxin-resolving on the migrating potentiality and associated gene expressive activities of nasopharyngeal carcinoma (NPC) cell line CNE2Z cells. Methods Firstly, different formulae derived from the same therapeutic principle of Qi-boosting toxin-resolving were composed only with their dosages changed but not with their composing kinds of herbs, i.e. the original formula (OF) and the modified one (MF). Herbal medicines-containing plasma (HMCP) were prepared routinely from rats by feeding method with these two formulae. Then, HMCP of these two formulae were cultured with CNE2Z cells in different concentrations respectively to detect their effects on migrating potentiality of these cells by scratching test. Immunohistochemical SABC assay was used to determine the expressive levels of nm23-H1 and c-myc in different groups of cells, and RT-PCR techniques were taken to confirm the transcribing activities of nm23-H1 mRNA and c-myc mRNA. Their interaction was evaluated by careful analysis on these data at last. Results TIt was shown from the experiment of scratching that the motility ability of targeting cells was significantly inhibited by the effect of OF and promoted by the effect of MF, inhibit potentiality of motility of CNE2, modified QBTRF promote potentiality of motility of CNE2. However, their effects on the expression of nm23-H1 and c-myc genes were rather complicated. OF showed no obvious effect on the expression of c-myc but giving rise a promoting effect on that of nm23-H1 protein (P<0.05), while MF induced promoting effect on the expressive activity of c-myc but inhibiting effect on that of nm23-H1 (P<0.05). However, results from RT-PCR assays indicated that the expressive levels of nm23-H1 mRNA were markedly elevated following the effects of OF and RF at higher and moderate dosages respectively(P<0.05), while lower dosages of both formulae had no obvious effects on their trascription(P>0.05). In contraction, MF produced a promoting effect on the expressive activity of c-myc mRNA at higher dosage, significantly higher than that of controlling group(P<0.05), but OF showed no marked effect on such an indicator at this concentration (P>0.05). Conclusion The original Qi-Boosting Toxin-Resolving formula gives rise an inhibitory effect on migrating potentiality of CNE2Z cells, while the modified Qi-Boosting Toxin-Resolving formula induces a kind of promoting effect on this potentiality of these cells, though only with the dosages changed without the herbal kinds changed in this formula. Such a kind of effect induced by MF may be associated with its special effects on the expression of nm23-H1 and c-myc genes and their interaction among them. It should be confirmed in following studies further. Moreover, the results from this work are also of significance for future study how to plan the dosage distribution rightly when composing a herbal formula.