Effect of Reduning Injection， Shenfu Injection Combined with Gefitinib on Proliferation， Stemness Characteristics and Metabolism of Non-small Cell Lung Cancer Cells

XIAO Zhenzhen; ZHU Yanjuan; YU Yaya; DING Lina; MA Changju; LEI Chenjing; LIU Yihong; CHANG Xuesong; CHEN Yadong; ZHANG Haibo

doi:10.13422/j.cnki.syfjx.20222421

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Effect of Reduning Injection， Shenfu Injection Combined with Gefitinib on Proliferation， Stemness Characteristics and Metabolism of Non-small Cell Lung Cancer Cells

更新时间：2023-08-22

- Effect of Reduning Injection， Shenfu Injection Combined with Gefitinib on Proliferation， Stemness Characteristics and Metabolism of Non-small Cell Lung Cancer Cells
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 18, Pages: 45-53(2023)
- 作者机构：
  
  1.广州中医药大学第二附属医院（广东省中医院），广州 510120
  2.广东省中医证候临床研究重点实验室，广州 510120
  3.粤港澳中医药与免疫疾病研究联合实验室，广州 510120
  4.广州中医药大学第二临床医学院，广州 510120
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20222421
  CLC： R22;R242;R2-031;R285.5
- Published：20 September 2023，
  
  Published Online：21 October 2022，
  
  Received：13 July 2022，
- 稿件说明：
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肖真真,朱燕娟,余娅娅等.热毒宁注射液、参附注射液联合吉非替尼对非小细胞肺癌细胞增殖、干性特征及代谢的影响[J].中国实验方剂学杂志,2023,29(18):45-53.

XIAO Zhenzhen,ZHU Yanjuan,YU Yaya,et al.Effect of Reduning Injection， Shenfu Injection Combined with Gefitinib on Proliferation， Stemness Characteristics and Metabolism of Non-small Cell Lung Cancer Cells[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(18):45-53.
肖真真,朱燕娟,余娅娅等.热毒宁注射液、参附注射液联合吉非替尼对非小细胞肺癌细胞增殖、干性特征及代谢的影响[J].中国实验方剂学杂志,2023,29(18):45-53. DOI： 10.13422/j.cnki.syfjx.20222421.

XIAO Zhenzhen,ZHU Yanjuan,YU Yaya,et al.Effect of Reduning Injection， Shenfu Injection Combined with Gefitinib on Proliferation， Stemness Characteristics and Metabolism of Non-small Cell Lung Cancer Cells[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(18):45-53. DOI： 10.13422/j.cnki.syfjx.20222421.

摘要

目的

选用以清热为主的热毒宁注射液，以温阳为主的参附注射液，对比观察清热、温阳两个相反治则中药联合吉非替尼对非小细胞肺癌细胞增殖、凋亡、干性特征及代谢的影响。

方法

选用不同的非小细胞肺癌细胞株，分别用吉非替尼（5、10、20 μmol·L

-1

）、热毒宁注射液（0.6%、0.9%）、参附注射液（0.6%、0.9%）、吉非替尼联合热毒宁注射液、吉非替尼联合参附注射液干预，采用细胞增殖与活性检测（CCK-8）法检测各组细胞的增殖情况；利用流式细胞术检测细胞的凋亡情况；运用实时荧光定量聚合酶链式反应（Real-time PCR）及蛋白免疫印迹法（Western blot）检测肺癌干细胞标志物性别决定区Y框蛋白2（Sox2）、乙酰脱氢酶家族成员A1 （ALDH1A1）mRNA及蛋白表达水平；采用飞秒激光无标记影像（FLI）技术检测肺癌细胞氧化还原比，运用能量代谢仪检测细胞的糖酵解水平。

结果

与空白组比较，热毒宁注射液能降低肺癌细胞存活率（

0.05），提高细胞凋亡率（

0.05），下调干细胞标志物Sox2和ALDH1A1 mRNA及蛋白表达水平（

0.05）、上调细胞的氧化还原比（

0.05），而参附注射液对细胞存活率、细胞凋亡率、Sox2和ALDH1A1的mRNA、蛋白表达水平及细胞氧化还原比的影响差异无统计学意义。与吉非替尼单药比较，热毒宁注射液联合吉非替尼可明显抑制肺癌细胞存活率（

0.05），促进肺癌细胞凋亡（

0.05），下调干细胞标志物Sox2和ALDH1A1 mRNA及蛋白表达水平（

0.05），上调细胞氧化还原比（

0.05），下调细胞糖酵解质子流出速率（

0.05），参附注射液联合吉非替尼对肺癌细胞的增殖、凋亡、干细胞标志物的表达水平、细胞氧化还原比及细胞糖酵解水平影响差异无统计学意义。

结论

清热中药热毒宁注射液可能通过调控肿瘤细胞的代谢，抑制其干性特征来增强吉非替尼对肺癌细胞的增殖抑制和促凋亡作用，温阳中药参附注射液对吉非替尼无明显的增效作用，表明表皮生长因子受体酪氨酸激酶抑制剂（EGFR-TKIs）应考虑与“清热”治则的中药联合应用。

Abstract

Objective

To compare and observe the effect of Reduning injection （mainly clearing heat）， Shenfu injection （mainly warming Yang） combined with gefitinib on the proliferation， apoptosis， stemness characteristics and metabolism of lung cancer cells.

Method

Different non-small cell lung cancer （NSCLC） cell lines were selected and intervened with gefitinib （5， 10， 20 μmol·L

-1

）， Reduning injection （0.6%， 0.9%）， Shenfu injection （0.6%， 0.9%）， gefitinib combined with Reduning injection， and gefitinib combined with Shenfu injection. Cell proliferation in each group was detected by cell counting kit-8 （CCK-8） assay， and cell apoptosis was detected by flow cytometry. The mRNA and protein expressions of lung cancer stem cell markers sex determining region Y-box 2 （Sox2） and aldehyde dehydrogenase family 1 member A1 （ALDH1A1） were determind by real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. The redox ratio of lung cancer cells was observed by femtosecond label-free imaging （FLI） and energy metabolism instrument was used to determine the glycolysis level in cells.

Result

Compared with the blank group， Reduning injection reduced the survival rate of lung cancer cells （

0.05）， increased the apoptosis rate （

0.05）， down-regulated the mRNA and protein expressions of Sox2 and ALDH1A1 （

0.05）， and up-regulated the redox ratio of cells （

0.05）， while Shenfu injection exerted no remarkable effect on the above indexes. In addition， compared with gefitinib alone， Reduning injection combined with gefitinib inhibited the survival rate of lung cancer cells （

0.05）， promoted the cell apoptosis （

0.05）， down-regulated the mRNA and protein expressions of Sox2 and ALDH1A1 （

0.05）， up-regulated the redox ratio of cells （

0.05）， and lowered the proton efflux rate of glycolysis （

0.05）， while Shenfu injection combined with gefitinib failed to affect these indexes of lung cancer cells significantly.

Conclusion

Reduning injection may inhibit stemness characteristics of tumor cells by regulating their metabolism to enhance the proliferation-inhibiting and pro-apoptotic effects of gefitinib on lung cancer cells， while Shenfu injection had no significant enhancing effect on gefitinib. This indicates that epidermal growth factor receptor tyrosine kinase inhibitors （EGFR-TKIs） should be used in combination with heat-clearing Chinese medicines.

关键词

热毒宁注射液参附注射液吉非替尼非小细胞肺癌（NSCLC）干性特征代谢

Keywords

Reduning injectionShenfu injectiongefitinibnon-small cell lung cancer （NSCLC）stemness characteristicsmetabolism

references

SUNG H，FERLAY J，SIEGEL R L，et al. Global cancer statistics 2020： GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries［J］. CA Cancer J Clin，2021，71（3）：209-249.

WU L L，KE L P，ZHANG Z S，et al. Development of EGFR TKIs and options to manage resistance of third-generation EGFR TKI osimertinib： Conventional ways and immune checkpoint inhibitors［J］.Front Oncol，2020， doi：10.3389/fonc.2020.602762http://dx.doi.org/10.3389/fonc.2020.602762.

ZHOU C C，WU Y L，CHEN G Y，et al. Erlotinib versus chemotherapy as first-line treatment for patients with advanced EGFR mutation-positive non-small-cell lung cancer （OPTIMAL，CTONG-0802）： A multicentre，open-label，randomised，phase 3 study［J］. Lancet Oncol，2011，12（8）：735-742.

FUKUOKA M，WU Y L，THONGPRASERT S，et al. Biomarker analyses and final overall survival results from a phase Ⅲ，randomized，open-label，first-line study of gefitinib versus carboplatin/paclitaxel in clinically selected patients with advanced non-small-cell lung cancer in Asia （IPASS）［J］. J Clin Oncol，2011，29（21）：2866-2874.

PROTO C，LO RUSSO G，CORRAO G，et al.Treatment in EGFR-mutated non-small cell lung cancer： How to block the receptor and overcome resistance mechanisms［J］.Tumori，2017，103（4）：325-337.

MOK T S，WU Y L，AHN M J，et al.Osimertinib or platinum-pemetrexed in EGFR T790M-positive lung cancer［J］.N Engl J Med，2017，376（7）：629-640.

HUANG T，SONG X，XU D，et al.Stem cell programs in cancer initiation，progression，and therapy resistance［J］.Theranostics，2020，10（19）：8721-8743.

李慧杰，胡熙文，齐元富，等.代谢重编程对攻毒治法靶向治疗肺癌干细胞的影响［J］.中国实验方剂学杂志，2020，26（8）：75-80.

程耿，周舰，张朝阳，等.姜黄素对前列腺癌干细胞维持及细胞糖代谢的影响［J］.沈阳药科大学学报，2022，39（6）：690-694，716.

刘璨，刘艾鑫，姜昌镐.木兰花碱通过ROS/KRAS/AMPK抑制结直肠癌sw480细胞干性特征和糖酵解［J］.中国免疫学杂志，2022，38（3）：344-347.

ZHU Y J，ZHANG H B，LIU L R，et al. Yin-cold or Yang-heat syndrome type of traditional Chinese medicine was associated with the epidermal growth factor receptor gene status in non-small cell lung cancer patients：Confirmation of a TCM concept［J］. Evid Based Complement Alternat Med，2017， doi：10.1155/2017/7063859http://dx.doi.org/10.1155/2017/7063859.

YU Y Y，ZHU Y J，ZOU Y，et al. Qing-Kai-Ling injection acts better than Shen-Fu injection in enhancing the antitumor effect of gefitinib in resistant non-small cell lung cancer models［J］. Evid Based Complement Alternat Med，2021， doi：10.1155/2021/9911935http://dx.doi.org/10.1155/2021/9911935.

ZHANG H，CHEN Y，CAO D，et al.Optical biopsy of laryngeal lesions using femtosecond multiphoton microscopy［J］.Biomed Opt Express，2021，12（3）：1308-1319.

SHI Y，AU J S，THONGPRASERT S，et al.A prospective，molecular epidemiology study of EGFR mutations in Asian patients with advanced non-small-cell lung cancer of adenocarcinoma histology （PIONEER）［J］.J Thorac Oncol，2014，9（2）：154-162.

CHANG W J，SUNG J S，LEE S Y，et al. The clinical significance of RAS，PIK3CA，and PTEN mutations in non-small cell lung cancer using cell-free DNA［J］. J Clin Med，2020，9（8）：2642.

ZHU Y J，QU X，ZHAN D D，et al. Specific gene co-variation acts better than number of concomitant altered genes in predicting EGFR-TKI efficacy in non-small-cell lung cancer［J］. Clin Lung Cancer，2021，22（1）： e98-e111.

朱燕娟，曲鑫，陈惠惠，等.非小细胞肺癌热毒证与基因突变及表皮生长因子酪氨酸激酶抑制剂疗效的相关性［J］.中华中医药杂志，2022，37（8）：4756-4760.

MORGILLO F，DELLA CORTE C M，FASANO M，et al. Mechanisms of resistance to EGFR-targeted drugs：Lung cancer［J］. ESMO Open，2016，1（3）：e000060.

ZHANG J，QU Z，YAO H，et al. An effective drug sensitizing agent increases gefitinib treatment by down regulating PI3K/Akt/mTOR pathway and up regulating autophagy in non-small cell lung cancer［J］. Biomed Pharmacother，2019，118：109169.

ZHU Y J，YING Z，CHEN H H，et al. Effect of Chinese medicine prescriptions of warming-Yang or cooling-heat combined with Gefitinib on H1650 cell line［J］. TMR Cancer，2018，1（1）：15-22.

ZHENG F，LI J，MA C，et al. Novel regulation of miR-34a-5p and HOTAIR by the combination of berberine and gefitinib leading to inhibition of EMT in human lung cancer［J］. J Cell Mol Med，2020，24（10）：5578-5592.

HENG W S，GOSENS R，KRUYT F.Lung cancer stem cells：Origin，features，maintenance mechanisms and therapeutic targeting［J］.Biochem Pharmacol，2019，160：121-133.

YAO W，WANG L，HUANG H，et al. All-trans retinoic acid reduces cancer stem cell-like cell-mediated resistance to gefitinib in NSCLC adenocarcinoma cells［J］. BMC Cancer，2020，20（1）：315.

SHIEN K，TOYOOKA S，YAMAMOTO H，et al.Acquired resistance to EGFR inhibitors is associated with a manifestation of stem cell-like properties in cancer cells［J］.Cancer Res，2013，73（10）：3051-3061.

BORA-SINGHAL N，MOHANKUMAR D，SAHA B，et al. Novel HDAC11 inhibitors suppress lung adenocarcinoma stem cell self-renewal and overcome drug resistance by suppressing Sox2［J］. Sci Rep，2020，10（1）：4722.

KOLENC O I，QUINN K P. Evaluating cell metabolism through autofluorescence imaging of NAD（P）H and FAD［J］. Antioxid Redox Signal，2019，30（6）：875-889.

VARONE A，XYLAS J，QUINN K P，et al.Endogenous two-photon fluorescence imaging elucidates metabolic changes related to enhanced glycolysis and glutamine consumption in precancerous epithelial tissues［J］.Cancer Res，2014，74（11）：3067-3075.

CODD A S，KANASEKI T，TORIGO T，et al.Cancer stem cells as targets for immunotherapy［J］.Immunology，2018，153（3）：304-314.

SONG I S，JEONG Y J，JEONG S H，et al. FOXM1-induced PRX3 regulates stemness and survival of colon cancer cells via maintenance of mitochondrial function［J］. Gastroenterology，2015，149：1006-1016.

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