Effect of Wuzhishan <italic style="font-style: italic">Callicarpa nudiflora</italic> in Hainan Province on Cisplatin-induced Apoptosis of Nasopharyngeal Carcinoma Cells and Mechanism： Based on Nrf2/ARE Pathway

Yi-hua SU; Yun-xin WANG; Jing-wen JIANG

doi:10.13422/j.cnki.syfjx.20220296

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Effect of Wuzhishan Callicarpa nudiflora in Hainan Province on Cisplatin-induced Apoptosis of Nasopharyngeal Carcinoma Cells and Mechanism： Based on Nrf2/ARE Pathway

更新时间：2021-12-14

- Effect of Wuzhishan Callicarpa nudiflora in Hainan Province on Cisplatin-induced Apoptosis of Nasopharyngeal Carcinoma Cells and Mechanism： Based on Nrf2/ARE Pathway
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 28, Issue 2, Pages: 131-138(2022)
- 作者机构：
  
  海南省中医院，海口 570203
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20220296
  CLC： R22;R242;R285.5;R2-031;R979.1
- Published：20 January 2022，
  
  Published Online：20 November 2021，
  
  Received：18 March 2021，
- 稿件说明：
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苏乙花,汪云鑫,姜靖雯.基于Nrf2/ARE信号通路探讨海南五指山裸花紫珠对鼻咽癌细胞顺铂诱导凋亡敏感性的作用及其机制[J].中国实验方剂学杂志,2022,28(02):131-138.

SU Yi-hua,WANG Yun-xin,JIANG Jing-wen.Effect of Wuzhishan Callicarpa nudiflora in Hainan Province on Cisplatin-induced Apoptosis of Nasopharyngeal Carcinoma Cells and Mechanism： Based on Nrf2/ARE Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(02):131-138.
苏乙花,汪云鑫,姜靖雯.基于Nrf2/ARE信号通路探讨海南五指山裸花紫珠对鼻咽癌细胞顺铂诱导凋亡敏感性的作用及其机制[J].中国实验方剂学杂志,2022,28(02):131-138. DOI： 10.13422/j.cnki.syfjx.20220296.

SU Yi-hua,WANG Yun-xin,JIANG Jing-wen.Effect of Wuzhishan Callicarpa nudiflora in Hainan Province on Cisplatin-induced Apoptosis of Nasopharyngeal Carcinoma Cells and Mechanism： Based on Nrf2/ARE Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(02):131-138. DOI： 10.13422/j.cnki.syfjx.20220296.

摘要

目的

探讨海南五指山裸花紫珠（LHZZ）对鼻咽癌（NPC）细胞顺铂（DDP）诱导凋亡敏感性的影响，并探究其作用机制。

方法

细胞增殖与活性检测（CCK-8）法检测不同浓度DDP（0，2，4，8，16，32 mg·L

-1

）和不同浓度海南五指山裸花紫珠（0，25，50，75，100 mg·L

-1

）处理后的细胞存活率。实验分组为空白组（正常HNE1细胞），DDP组（8 mg·L

-1

，24 h），LHZZ组（50 mg·L

-1

，24 h），DDP+LHZZ组（8 mg·L

-1

DDP+50 mg·L

-1

LHZZ，24 h），DDP+LHZZ+核因子E2相关因子2（Nrf2）激活剂莱菔硫烷（SFN）组（8 mg·L

-1

DDP+50 mg·L

-1

LHZZ，24 h；然后10 μmol·L

-1

SFN，24 h），CCK-8法检测细胞存活率，集落克隆形成实验检测细胞集落形成能力，流式细胞术和原位末端标记法（TUNEL）染色检测细胞凋亡情况，荧光探针2'，7'-二氯荧光黄双乙酸盐（DCFH-DA）检测培养细胞活性氧（ROS）水平，蛋白免疫印迹法（Western blot）检测细胞内凋亡相关蛋白B细胞淋巴瘤-2（Bcl-2），Bcl-2相关X 蛋白（Bax）和半胱氨酸天冬氨酸蛋白水解酶-3（Caspase-3）表达，实时荧光定量聚合酶链式反应（Real-time PCR）检测细胞内Nrf2，抗氧化反应元件（ARE） mRNA表达。

结果

与空白组比较，经过不同浓度DDP和不同浓度LHZZ处理后，细胞存活率均明显下降（

0.05）；与DDP组和LHZZ组比较，DDP+LHZZ组细胞存活率下降，细胞集落克隆形成数目减少，细胞凋亡水平升高，Bcl-2蛋白表达水平下调，Bax和Caspase-3蛋白表达水平则上调，差异具有统计学意义（

0.05）；而在DDP+LHZZ组中加入SFN激活Nrf2/ARE信号通路，上述效果均受到抑制（

0.05）。此外，与空白组比较，LHZZ组细胞内ROS水平下调（

0.05）；与DDP组比较，DDP+LHZZ组ROS水平明显下调（

0.05）；与DDP+LHZZ组比较，DDP+LHZZ+SFN组细胞ROS水平明显上调（

0.05）。

结论

LHZZ可增加DDP诱导的鼻咽癌细胞凋亡的敏感性，其机制可能与阻断Nrf2/ARE信号通路并抑制ROS水平相关。

Abstract

Objective

To explore the effect of Wuzhishan

Callicarpa nudiflora

（LHZZ） on the sensitivity of nasopharyngeal carcinoma （NPC） cells to cisplatin （DDP） and the mechanism.

Method

Cell counting kit-8 （CCK-8） assay was used to detect the survival rate of NPC HNE1 cells after treatment with different concentration of DDP （0，2，4，8，16，32 mg·L

-1

） and different concentration of LHZZ （0，25，50，75，100 mg·L

-1

）. The following groups were designed： control group （normal HNE1 cells），DDP group （8 mg·L

-1

DDP，24 h），LHZZ group （50 mg·L

-1

LHZZ，24 h），DDP + LHZZ group （8 mg·L

-1

DDP + 50 mg·L

-1

LHZZ，24 h），DDP + LHZZ + nuclear factor erythroid 2-related factor 2 （Nrf2） activator sulforaphane （SFN） group （8 mg·L

-1

DDP + 50 mg·L

-1

LHZZ，24 h， 10 μmol·L

-1

SFN，24 h）. Then CCK-8 assay was employed to examine cell survival rate，colony formation test the colony-forming ability，flow cytometry and in situ terminal end-labeling（TUNEL） staining cell apoptosis，fluorescent probe 2'，7'-dichlorofluorescin diacetate （DCFH-DA） the level of reactive oxygen species （ROS） in cultured cells，Western blot the expression of apoptosis-related proteins in cells，such as B-cell lymphoma/leukemia-2 （Bcl-2），Bcl-2 associated X protein （Bax），and cysteinyl aspartate specific proteinase-3 （Caspase-3），and Real-time quantitative polymerase chain reaction （Real-time PCR） the expression of Nrf2 and antioxidant response element （ARE） mRNA in cells.

Result

The survival rates of cells treated with different concentration of DDP and different concentration of LHZZ decreased compared with that in the control group （

0.05）. Compared with the DDP group and the LHZZ group，DDP + LHZZ group demonstrated decrease in cell survival rate，number of cell colonies，and Bcl-2 level，and increase in the apoptosis level and the expression of Bax and Caspase-3 （

0.05）. However，after the addition of SFN，the Nrf2/ARE signaling pathway was activated and the above variation was inhibited （

0.05）. In addition，the level of intracellular ROS in the LHZZ group was lower than that in the control group （

0.05） and the level in the DDP + LHZZ group was lower than that in the DDP group （

0.05）. Moreover，the ROS level in the DDP + LHZZ + SFN group was higher than that in the DDP+LHZZ group （

0.05）.

Conclusion

LHZZ can enhance the sensitivity of DDP-induced NPC apoptosis，possibly by blocking the Nrf2/ARE signaling pathway and inhibiting the level of ROS.

关键词

鼻咽癌顺铂海南五指山裸花紫珠凋亡敏感性

Keywords

nasopharyngeal carcinomacisplatinWuzhishan Callicarpa nudiflora in Hainan provinceapoptosissensitivity

references

CHEN Y P，CHAN A T C，LE Q T，et al. Nasopharyngeal carcinoma［J］. Lancet，2019，394（10192）：64.

NG W T，TUNG S Y，LEE V，et al. Concurrent-adjuvant chemoradiation therapy for stage Ⅲ-ⅣB nasopharyngeal carcinoma-exploration for achieving optimal 10-year therapeutic ratio［J］. Int J Radiat Oncol Biol Phys，2018，101（5）：78.

MOON S H，CHO K H，LEE C G，et al. IMRT vs. 2D-radiotherapy or 3D-conformal radiotherapy of nasopharyngeal carcinoma ：survival outcome in a Korean multi-institutional retrospective study （KROG 11-06）［J］. Strahlenther Onkol，2016，192（6）：377-385.

LEE A W M，NG W T，CHAN J Y W，et al. Management of locally recurrent nasopharyngeal carcinoma［J］. Cancer Treat Rev，2019，79：101890.

LIANG S B，CHEN L S，YANG X L，et al. Influence of tumor necrosis on treatment sensitivity and long-term survival in nasopharyngeal carcinoma［J］.Radiother Oncol，2020，18：219-225.

张力军，陈曦，冯世，等. 基于系统药理学探索裸花紫珠有效成分的作用机制［J］. 药物评价研究，2018，41（10）：1759-1769.

FU H Z，MA Y Y，MA S C，et al. Two new iridoid glycosides from Callicarpa nudiflora［J］ .J Asian Nat Prod Res，2020，22（3）：264-270.

占丽丽，叶先文，张敏，等. 紫珠属植物化学成分及药理活性研究进展［J］. 江西中医药，2020，51（8）：66-73.

WU Y S，SHI L，LIU X G，et al. Chemical profiling of Callicarpa nudiflora and its effective compounds identification by compound-target network analysis［J］. J Pharm Biomed Anal，2020，182：113110.

陈学武，姜靖雯，张永杰.海南五指山裸花紫珠通过IL-6/STAT3信号通路对结肠癌体外耐药的逆转作用及其机制研究［J］.中药材，2020，43（9）：2269-2273.

张仲，罗贤强，周金娜，等. 正交试验优化裸花紫珠水提取工艺及外用止血作用的评估［J］. 中国现代中药，2018，20（6）：734-737.

LAM W K J，CHAN J Y K. Recent advances in the management of nasopharyngeal carcinoma［J］.F1000Res，2018，dio：10.12688/f1000research 15066.1.

黄岚晖，韦小娜，刘俊波.化疗联合放射治疗在局部晚期鼻咽癌中的应用效果［J］.微创医学，2020，15（3）：335-337.

ZHANG Y，CHEN L，HU G Q，et al. Gemcitabine and cisplatin induction chemotherapy in nasopharyngeal carcinoma［J］. N Engl J Med，2019，381（12）：1124-1135.

VERMORKEN J B，STÖHLMACHER-WILLIAMS J，DAVIDENKO I，et al. Cisplatin and fluorouracil with or without panitumumab in patients with recurrent or metastatic squamous-cell carcinoma of the head and neck （SPECTRUM）：an open-label phase 3 randomised trial［J］. Lancet Oncol，2013，14（8）：697-710.

SHELTON P，JAISWAL A K. The transcription factor NF-E2-related factor 2 （nrf2）：a protooncogene？［J］. FASEB J，2013，27（2）：414-423.

GONG M，LI Y，YE X，et al. Loss-of-function mutations in KEAP1 drive lung cancer progression via KEAP1/NRF2 pathway activation［J］. Cell Commun Signal，2020，18（1）：98.

TIAN Y，WU K，LIU Q，et al. Modification of platinum sensitivity by KEAP1/NRF2 signals in non-small cell lung cancer［J］. J Hematol Oncol，2016，9（1）：83.

TAGUCHI K，MOTOHASHI H，YAMAMOTO M. Molecular mechanisms of the Keap1-Nrf2 pathway in stress response and cancer evolution［J］. Gene Cell，2011，16（2）：123-140.

YAMAZAKI H，TANJI K，WAKABAYASHI K，et al. Role of the Keap1/Nrf2 pathway in neurode⁃generative diseases［J］. Pathol Int，2015，65（5）：210-219.

JI L，LI H，GAO P，et al. Nrf2 pathway regulates multidrug-resistance-associated Protein 1 in small cell lung cancer［J］. PLoS One，2013，8（5）：e63404.

FOUZDER C，MUKHUTY A，MUKHERJEE S，et al. Trigonelline inhibits Nrf2 via EGFR signalling pathway and augments efficacy of Cisplatin and Etoposide in NSCLC cells［J］. Toxicol In Vitro，2021，70：105038.

CHEN B，LU Y，CHEN Y，et al. The role of Nrf2 in oxidative stress-induced endothelial injuries［J］. J Endocrinol，2015，225（3）：R83-R99.

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Effect of Wuzhishan Callicarpa nudiflora in Hainan Province on Cisplatin-induced Apoptosis of Nasopharyngeal Carcinoma Cells and Mechanism： Based on Nrf2/ARE Pathway

DOI：10.13422/j.cnki.syfjx.20220296

摘要

Abstract

关键词

Keywords

references