| 引用本文: | 李亚洲,周燕园,白红妍,董哲文,林雯,付红红,董玲,廖燕梅,吕良.青天葵甲醇提取物通过ERK信号通路诱导鼻咽癌CNE-2细胞凋亡[J].中国现代应用药学,2021,38(16):1928-1933. |
| LI Yazhou,ZHOU Yanyuan,BAI Hongyan,DONG Zhewen,LIN Wen,FU Honghong,DONG Ling,LIAO Yanmei,LYU Liang.Methanol Extract of Nervilia Fordii Induces Apoptosis of Nasopharyngeal Carcinoma CNE-2 Cells Through ERK Signaling Pathway[J].Chin J Mod Appl Pharm(中国现代应用药学),2021,38(16):1928-1933. |
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| 青天葵甲醇提取物通过ERK信号通路诱导鼻咽癌CNE-2细胞凋亡 |
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李亚洲1, 周燕园1, 白红妍1, 董哲文1, 林雯1, 付红红1, 董玲1, 廖燕梅1, 吕良1,2
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1.桂林医学院药学院, 广西 桂林 541004;2.广西医科大学广西区域性高发肿瘤早期防治研究重点实验室, 南宁 530000
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| 摘要: |
| 目的 研究青天葵甲醇提取物（Nervilia fordii methanol extracts，NFME）体外对鼻咽癌CNE-2细胞的凋亡作用及其作用机制。方法 MTT法检测不同浓度（0，0.25，0.5，1，2，3 mg·mL-1）的NFME处理CNE-2细胞24，48 h对CNE-2细胞生长抑制率的影响；克隆原形成试验观察NFME对CNE-2细胞克隆形成率的影响；Hoechst凋亡染色观察NFME对CNE-2细胞凋亡的影响；Western blotting测定NFME作用下caspase-3、ERK1/2和c-Raf蛋白磷酸化水平的变化。结果 MTT试验结果显示，与对照组相比，在给药24 h时0.5 mg·mL-1的NFME就能抑制CNE-2细胞的增殖（P<0.05），抑制率为12.64%。而在给药48 h时0.25 mg·mL-1的NFME就能抑制CNE-2细胞增殖（P<0.05），抑制率为22.43%；克隆原形成能力试验表明，0.25 mg·mL-1的NFME能够抑制CNE-2细胞集落的形成（P<0.05）；Hoechst33258凋亡染色观察到0.25 mg·mL-1的NFME作用24 h能够观察到CNE-2细胞发生凋亡（P<0.05），凋亡率达到17.91%；Western blotting结果表明0.5 mg·mL-1的NFME能使CNE-2细胞中caspase-3发生剪切，随着给药浓度增加其剪切作用越明显（P<0.01），同时0.5 mg·mL-1的NFME能够降低CNE-2细胞中ERK1/2和c-Raf蛋白的磷酸化水平（P<0.05）。结论 青天葵甲醇提取物能抑制鼻咽癌CNE-2细胞的增殖并诱导其发生凋亡，其作用机制可能与抑制ERK信号通路有关。 |
| 关键词: 青天葵甲醇提取物 鼻咽癌 CNE-2 ERK 凋亡 |
| DOI：10.13748/j.cnki.issn1007-7693.2021.16.002 |
| 分类号:R285.5 |
| 基金项目:国家自然科学基金项目（81860767）；广西区域性高发肿瘤早期防治研究重点实验室开放课题（GKE2018-KF04）；广西自然科学基金资助项目（2018GXNSFAA281104）；广西高等学校千名中青年骨干教师培育计划资助项目；桂林医学院中青年教职工科研能力提升项目（2018glmcy010） |
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| Methanol Extract of Nervilia Fordii Induces Apoptosis of Nasopharyngeal Carcinoma CNE-2 Cells Through ERK Signaling Pathway |
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LI Yazhou1, ZHOU Yanyuan1, BAI Hongyan1, DONG Zhewen1, LIN Wen1, FU Honghong1, DONG Ling1, LIAO Yanmei1, LYU Liang1,2
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1.College of Pharmacy, Guilin Medical University, Guilin 541004, China;2.Guangxi Key Laboratory of Early Prevention and Treatment for Regional High Frequency Tumor, Guangxi Medical University, Nanning 530000, China
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| Abstract: |
| OBJECTIVE To investigate the apoptosis effect and mechanism of Nervilia fordii methanol extracts(NFME) on nasopharyngeal carcinoma CNE-2 cells in vitro. METHODS The growth inhibition rate of CNE-2 cells treated with different concentrations of NFME(0, 0.25, 0.5, 1, 2, 3 mg·mL-1) for 24, 48 h were detected by MTT. The effect of NFME on CNE-2 cell clone formation rate was observed by clonal test. The effect of NFME on CNE-2 cell apoptosis was observed by Hoechst apoptosis staining. The changes of phosphorylation level of caspase-3, ERK1/2 and c-Raf were detected by Western blotting. RESULTS The results of the MTT experiment showed that, compared with the control group, 0.5 mg·mL-1 of NFME could inhibit the proliferation of CNE-2 cells at 24 h(P<0.05), and the inhibition rate reached 12.64%. At 48 h, 0.25 mg·mL-1 of NFME could inhibit the proliferation of CNE-2 cells(P<0.05), and the inhibition rate reached 22.43%. The experiment of clonal ability showed that 0.25 mg·mL-1 of NFME could inhibit CNE-2 formation of cell colonies(P<0.05). Hoechst33258 apoptosis staining observed that 0.25 mg·mL-1 of NFME induced apoptosis in CNE-2 cells for 24 h(P<0.05), and the apoptosis rate reached 17.91%. The results of Western blotting showed that 0.5 mg·mL-1 of NFME caused caspase-3 to shear in CNE-2 cells, and the shear effect became more pronounced as the concentration of drug was increased(P<0.01). Meanwhile, NFME could cut down the phosphorylation level of ERK1/2 and c-Raf in CNE-2 cells at 0.5 mg·mL-1(P<0.05). CONCLUSION NFME can inhibit nasopharyngeal carcinoma CNE-2 cells and induce apoptosis, its mechanism may be related to the inhibition of ERK signaling pathway. |
| Key words: Nervilia Fordii methanol extracts nasopharyngeal carcinoma CNE-2 ERK apoptosis |
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