甘肃中医药大学药学院,兰州 730000
Received:30 April 2023,
Revised:2023-06-20,
Accepted:13 July 2023,
Published:28 October 2023
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张尚龙,张楠,连小龙,等.甘草内生菌联合顺铂对A549细胞的增殖及凋亡的影响[J].重庆医科大学学报,2023,48(10):1173-1179.
Zhang Shanglong,Zhang Nan,Lian Xiaolong,et al.Effect of Glycyrrhiza uralensis Fisch. endophytes combined with cisplatin on proliferation and apoptosis of A549 cells[J].Journal of Chongqing Medical University,2023,48(10):1173-1179.
张尚龙,张楠,连小龙,等.甘草内生菌联合顺铂对A549细胞的增殖及凋亡的影响[J].重庆医科大学学报,2023,48(10):1173-1179. DOI: 10.13406/j.cnki.cyxb.003356.
Zhang Shanglong,Zhang Nan,Lian Xiaolong,et al.Effect of Glycyrrhiza uralensis Fisch. endophytes combined with cisplatin on proliferation and apoptosis of A549 cells[J].Journal of Chongqing Medical University,2023,48(10):1173-1179. DOI: 10.13406/j.cnki.cyxb.003356.
目的
2
探讨甘草内生菌JTZB55联合顺铂对人肺癌A549细胞增殖和凋亡的影响及其可能机制。
方法
2
MTT法检测JTZB55或/和顺铂对A549细胞存活率的影响,根据协同指数筛选出后续实验浓度,将A549细胞分为正常对照组、2 μg/mL顺铂组、800 μg/mL JTZB55组、2 μg/mL顺铂+800 μg/mL JTZB55组;流式细胞术检测各组药物对A549细胞凋亡的影响;Western blot法检测各组药物处理后细胞线粒体凋亡及内质网应激途径相关蛋白的表达水平。
结果
2
MTT结果显示,顺铂及JTZB55作用于A549细胞后,细胞存活率显著降低;与顺铂组相比,联合组细胞存活率显著降低(
P
<
0.05),且800 μg/mL JTZB55与2 μg/mL顺铂联用的协同指数最高;流式细胞术结果显示,JTZB55及顺铂均能促进A549细胞凋亡(
P
<
0.01),两药联合组细胞凋亡率显著增高(
P
<
0.01);Western blot实验结果表明,顺铂联合JTZB55能显著降低B细
胞淋巴瘤/白血病-2蛋白(B cell lymphoma/leukemia-2,Bcl-2)蛋白的表达,上调Bcl-2相关X蛋白(Bcl-2 associated X,Bax)、细胞色素C(Cytochrome C,CytC)、半胱氨酸天冬氨酸蛋白酶-3(cysteine aspartate protease-3,Caspase-3)、半胱氨酸天冬氨酸蛋白酶-8(cysteine aspartate protease-8,Caspase-8)、半胱氨酸天冬氨酸蛋白酶-9(cysteine aspartate protease-9,Caspase-9)、葡萄糖调节反应蛋白78(Glucose regulated protein 78,GRP78)、CCAAT-增强子结合蛋白同源蛋白(CCAAT/enhancer-binding protein-homologus protein,CHOP)蛋白的表达(
P
<
0.05)。
结论
2
JTZB55可以增强顺铂对A549细胞的增殖抑制作用,促进A549细胞的凋亡,其机制可能与下调Bcl-2和上调Bax、CytC、Caspase-3、Caspase-8、Caspase-9、GRP78、CHOP蛋白的表达相关。
Objective
2
To investigate the effect of
Glycyrrhiza uralensis Fisch.
endophyte JTZB55 combined with cisplatin on proliferation and apoptosis of human lung cancer A549 cells and the possible mechanism.
Methods
2
MTT assay was used to determine the effect of JTZB55 and/or cisplatin on the survival rate of A549 cells. According to the synergistic index,the subsequent experimental concentration was screened out,and A549 cells were divided into normal control group,2 μg/mL cisplatin group,800 μg/mL JTZB55 group,and 2 μg/mL cisplatin + 800 μg/mL JTZB55 group. Flow cytometry was used to determine the effect of drugs in each group on the apoptosis of A549 cells. Western blot was used to measure the expression levels of mitochondrial apoptosis and endoplasmic reticulum stress pathway-related proteins in cells treated with drugs in each group.
Results
2
MTT results showed that the survival rate of A549 cells was decreased significantly after cisplatin and JTZB55 were administrated. Compared with the cisplatin group,the cell survival rate in the combination group was significantly decreased(
P
<
0.05),and the synergistic index of 800 μg/mL JTZB55 combined with 2 μg/mL cisplatin was the highest. The results of flow cytometry showed that both JTZB55 and cisplatin could
promote the apoptosis of A549 cells(
P
<
0.01),and the apoptosis rate of A549 cells was significantly increased in the combination group(
P
<
0.01). Western blot results showed that cisplatin combined with JTZB55 could significantly decrease the expression of B cell lymphoma/leukemia-2(Bcl-2) protein and up-regulate the expression of Bcl-2 associated X(Bax),cytochrome C(CytC),cysteine aspartate protease-3(Caspase-3),cysteine aspartate protease-8(Caspase-8),cysteine aspartate protease-9(Caspase-9),glucose regulated protein 78(GRP78),and CCAAT/enhancer-binding protein-homologous protein(CHOP)(
P
<
0.05).
Conclusion
2
JTZB55 can enhance the inhibitory effect of cisplatin on the proliferation of A549 cells and promote the apoptosis of A549 cells. The mechanism may be related to the down-regulation of Bcl-2 and up-regulation of Bax,CytC,Caspase-3,Caspase-8,Caspase-9,GRP78,and CHOP.
Gómez-Ruiz S , Maksimović-Ivanić D , Mijatović S , et al . On the discovery,biological effects,and use of Cisplatin and metallocenes in anticancer chemotherapy [J ] . Bioinorg Chem Appl , 2012 , 2012 : 140284 .
Jamieson ER , Lippard SJ . Structure,recognition,and processing of cisplatin-DNA adducts [J ] . Chem Rev , 1999 , 99 ( 9 ): 2467 - 2498 .
Karadeniz A , Simsek N , Karakus E , et al . Royal jelly modulates oxidative stress and apoptosis in liver and kidneys of rats treated with cisplatin [J ] . Oxid Med Cell Longev , 2011 , 2011 : 981793 .
Jakupec MA , Galanski MS , Keppler BK . Tumour-inhibiting platinum complexes:state of the art and future perspectives [J ] . Rev Physiol Biochem Pharmacol , 2003 , 146 : 1 - 54 .
Rabik CA , Dolan ME . Molecular mechanisms of resistance and toxicity associated with platinating agents [J ] . Cancer Treat Rev , 2007 , 33 ( 1 ): 9 - 23 .
Gao Y , Chu SF , Shao QH , et al . Antioxidant activities of ginsenoside Rg1 against cisplatin-induced hepatic injury through Nrf2 signaling pathway in mice [J ] . Free Radic Res , 2017 , 51 ( 1 ): 1 - 13 .
Isoda K , Nozawa T , Taira Y , et al . Effects of surface charge and palladium on hepatic and kidney injury induced by polystyrene nanoparticles co-administered to mice with paraquat and cisplatin [J ] . Pharmazie , 2018 , 73 ( 3 ): 165 - 168 .
K V A , Madhana RM , Kasala ER , et al . Morin hydrate mitigates cisplatin-induced renal and hepatic injury by impeding oxidative/nitrosative stress and inflammation in mice [J ] . J Biochem Mol Toxicol , 2016 , 30 ( 12 ): 571 - 579 .
Ekinci Akdemir FN , Albayrak M , Çalik M , et al . The protective effects of p-coumaric acid on acute liver and kidney damages induced by cisplatin [J ] . Biomedicines , 2017 , 5 ( 2 ): 18 .
Boroja T , Katanić J , Rosić G , et al . Summer savory( Satureja hortensis L. ) extract:Phytochemical profile and modulation of cisplatin-induced liver,renal and testicular toxicity [J ] . Food Chem Toxicol , 2018 , 118 : 252 - 263 .
Bunel V , Antoine MH , Nortier J , et al . Nephroprotective effects of ferulic acid,Z-ligustilide and E-ligustilide isolated from Angelica sinensis against cisplatin toxicity in vitro [J ] . Toxicol Vitro , 2015 , 29 ( 3 ): 458 - 467 .
Kilic K , Sakat MS , Akdemir FNE , et al . Protective effect of Gallic acid against cisplatin-induced ototoxicity in rats [J ] . Braz J Otorhinolaryngol , 2019 , 85 ( 3 ): 267 - 274 .
Sherif IO , Abdel-Aziz A , Sarhan OM . Cisplatin-induced testicular toxicity in rats:the protective effect of arjunolic acid [J ] . J Biochem Mol Toxicol , 2014 , 28 ( 11 ): 515 - 521 .
李葆林 , 麻景梅 , 田宇柔 , 等 . 甘草中新发现化学成分和药理作用的研究进展 [J ] . 中草药 , 2021 , 52 ( 8 ): 2438 - 2448 .
Li BL , Ma JM , Tian YR , et al . Research progress on newly discovered chemical constituents and pharmacological effects of Glycyrrhizae Radix et Rhizoma [J ] . Chin Tradit Herb Drugs , 2021 , 52 ( 8 ): 2438 - 2448 .
张云云 , 吴开祥 , 孙雯雯 , 等 . 甘草查尔酮A促进MCF-7乳腺癌细胞TRAIL 敏感性的机制研究 [J ] . 中药材 , 2022 , 45 ( 4 ): 974 - 977 .
Zhang YY , Wu KX , Sun WW , et al . Mechanism of Glycyrrhiza chalcone A promoting TRAIL sensitivity in MCF-7 breast cancer cells [J ] . J Chin Med Mater , 2022 , 45 ( 4 ): 974 - 977 .
王 勇 , 黄大兵 , 沈夏波 , 等 . 甘草素调控WIG-1基因对肺癌细胞增殖、细胞凋亡以及放疗敏感性影响的研究 [J ] . 中国免疫学杂志 , 2020 , 36 ( 10 ): 1194 - 1200 .
Wang Y , Huang DB , Shen XB , et al . Liquiritigenin effects on proliferation,apoptosis and radio-sensitivity of lung carcinoma cells by regulating WIG-1 gene [J ] . Chin J Immunol , 2020 , 36 ( 10 ): 1194 - 1200 .
郝文龙 , 荔志云 . 异甘草素在脑胶质瘤治疗中的作用与进展 [J ] . 西北国防医学杂志 , 2019 , 40 ( 4 ): 255 - 259 .
Hao WL , Li ZY . Effect and progress of isoliquiritigenin in the treatment of glioma [J ] . Med J Natl Defending Forces Northwest China , 2019 , 40 ( 4 ): 255 - 259 .
张九成 , 陈卫东 , 邹 宁 . 甘草查尔酮A调控人结肠癌细胞增殖与凋亡的分子机制研究 [J ] . 中国免疫学杂志 , 2019 , 35 ( 5 ): 549 - 554 .
Zhang JC , Chen WD , Zou N . Molecular mechanism of Licochalcone A regulating proliferation and apoptosis of human colon cancer cells [J ] . Chin J Immunol , 2019 , 35 ( 5 ): 549 - 554 .
闫 潇 , 马 骏 , 张楠 , 等 . 马钱子总生物碱致肾毒性模型建立及甘草内生菌代谢产物减毒作用对比 [J ] . 中药药理与临床 , 2022 , 38 ( 4 ): 121 - 126 .
Yan X , Ma J , Zhang N , et al . Establishment of nephrotoxicity model induced by total alkaloids of strychni semen and comparison on attenuation effects of endophytes metabolites from glycyrrhizae radix et rhizoma [J ] . Pharmacol Clin Chin Mater Med , 2022 , 38 ( 4 ): 121 - 126 .
刘智丹 . 姜黄素联合顺铂诱导腺样囊性癌ACC-M细胞凋亡及对内质网应激相关蛋白表达的影响 [D ] . 遵义 : 遵义医科大学 , 2021 .
Liu ZD . Effect of curcumin combined with cisplatin on apoptosis and epression of endoplasmic reticulum stress related proteins in ACC-M cells of adenoid cystic carcinoma [D ] . Zunyi : Zunyi Med Univ , 2021 .
连小龙 , 令 颖 , 张尚龙 , 等 . 甘草有效成分抗肿瘤作用机制及联合抗肿瘤研究进展 [J ] . 中国实验方剂学杂志 , 2022 , 28 ( 11 ): 238 - 245 .
Lian XL , Ling Y , Zhang SL , et al . Anti-tumor mechanism of active ingredients of glycyrrhizae Radix et rhizoma and its combined use in anti-tumor treatment:a review [J ] . Chin J Exp Tradit Med Formulae , 2022 , 28 ( 11 ): 238 - 245 .
Iizaka T , Tsuji M , Oyamada H , et al . Interaction between caspase-8 activation and endoplasmic reticulum stress in glycochenodeoxycholic acid-induced apoptotic HepG2 cells [J ] . Toxicology , 2007 , 241 ( 3 ): 146 - 156 .
Zhao H , Yenari MA , Cheng DY , et al . Bcl-2 overexpression protects against neuron loss within the ischemic margin following experimental stroke and inhibits cytochrome c translocation and caspase-3 activity [J ] . J Neurochem , 2003 , 85 ( 4 ): 1026 - 1036 .
张丽惠 , 张琬莹 , 张志鹏 , 等 . 复方苦参注射液联合表柔比星对膀胱癌细胞凋亡的影响 [J ] . 中药药理与临床 , 2020 , 36 ( 3 ): 203 - 207 .
Zhang LH , Zhang WY , Zhang ZP , et al . Effect of compound Kushen injection combined with epirubicin on apoptosis of bladder cancer cells [J ] . Pharmacol Clin Chin Mater Med , 2020 , 36 ( 3 ): 203 - 207 .
王 娜 , 张 霞 . 甘草甜素调节U14宫颈癌小鼠免疫功能及诱导凋亡的作用 [J ] . 食品工业科技 , 2021 , 42 ( 23 ): 370 - 376 .
Wang N , Zhang X . Effect of glycyrrhizin on immune function and apoptosis of U 14 Cervical cancer mice [J ] . Sci Technol Food Ind , 2021 , 42 ( 23 ): 370 - 376 .
Li JJ , Tang F , Li RK , et al . Dietary compound glycyrrhetinic acid suppresses tumor angiogenesis and growth by modulating antiangiogenic and proapoptotic pathways in vitro and in vivo [J ] . J Nutr Biochem , 2020 , 77 : 108268 .
Li YM , Guo YS , Tang JA , et al . New insights into the roles of CHOP-induced apoptosis in ER stress [J ] . Acta Biochim Biophys Sin , 2014 , 46 ( 8 ): 629 - 640 .
Limonta P , Moretti RM , Marzagalli M , et al . Role of endoplasmic reticulum stress in the anticancer activity of natural compounds [J ] . Int J Mol Sci , 2019 , 20 ( 4 ): 961 .
袁永贵 , 张夏炎 , 朱晓俊 , 等 . 甘草查尔酮A对人乳腺癌MDA-MB-231细胞凋亡的影响 [J ] . 中国实验方剂学杂志 , 2021 , 27 ( 20 ): 95 - 100 .
Yuan YG , Zhang XY , Zhu XJ , et al . Effect of licochalcone A on apoptosis in human breast cancer MDA-MB-231 cells [J ] . Chin J Exp Tradit Med Formulae , 2021 , 27 ( 20 ): 95 - 100 .
张冉浠 , 周 莉 , 陆安伟 , 等 . GRP78在不同亚型宫颈癌细胞株中表达情况及对顺铂敏感性影响 [J ] . 肿瘤预防与治疗 , 2020 , 33 ( 5 ): 389 - 394 .
Zhang RX , Zhou L , Lu AW , et al . Expression of GRP78 and its effect on cisplatin sensitivity in different subtypes of cervical cancer cells [J ] . J Cancer Contr Treat , 2020 , 33 ( 5 ): 389 - 394 .
Zhang L , Cheng X , Xu SC , et al . Curcumin induces endoplasmic reticulum stress-associated apoptosis in human papillary thyroid carcinoma BCPAP cells via disruption of intracellular calcium homeostasis [J ] . Medicine , 2018 , 97 ( 24 ): e11095 .
Chang YJ , Huang CY , Hung CS , et al . GRP78 mediates the therapeutic efficacy of curcumin on colon cancer [J ] . Tumor Biol , 2015 , 36 ( 2 ): 633 - 641 .
张美兰 , 沈哲式 , 张英哲 , 等 . 姜黄素诱导人肝癌BEL-7404细胞凋亡的内质网应激作用的研究 [J ] . 吉林医学 , 2021 , 42 ( 1 ): 5 - 8 .
Zhang ML , Shen ZS , Zhang YZ , et al . Curcumin induces endoplasmic reticulum stress- apoptosis in human liver cancer BEL-7404 cell [J ] . Jilin Med J , 2021 , 42 ( 1 ): 5 - 8 .
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