Honokiol combined with curcumin sensitizes multidrug‑resistant human lung adenocarcinoma A549/DDP cells to cisplatin

Qi,Mingming; Chen,Xiaojin; Bian,Liqun; Zhang,Han; Ma,Jian

doi:10.3892/etm.2021.10736

Honokiol combined with curcumin sensitizes multidrug‑resistant human lung adenocarcinoma A549/DDP cells to cisplatin

Authors:
- Mingming Qi
- Xiaojin Chen
- Liqun Bian
- Han Zhang
- Jian Ma
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Affiliations: Department of Febrile Diseases, School of Chinese Medicine, Nanjing, Jiangsu 210023, P.R. China, Hanlin College, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, P.R. China, Digestive Department, Xiyuan Hospital, China Academy of Chinese Medicine Sciences, Beijing 100091, P.R. China
Published online on: September 16, 2021 https://doi.org/10.3892/etm.2021.10736
Article Number: 1301

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Abstract

The aim of the present study was to discuss the effects and underlying mechanisms of honokiol (HNK) and/or curcumin (CUR) in sensitization of multidrug‑resistant human lung adenocarcinoma A549/DDP cells to cisplatin (DDP). An MTS assay was performed to detect the cytotoxicity of HNK, CUR and DDP in A549 and A549/DDP cells and compare their sensitivity. The A549/DDP cells were then divided into 8 groups: Control, HNK, CUR, DDP, HNK + CUR, HNK + DDP, CUR + DDP and HNK + CUR + DDP. Cell proliferation was measured by MTS assay and colony formation assay, cell apoptosis was detected by flow cytometry, cell invasion was evaluated by Transwell assay and cell migration was determined by a wound healing assay. In order to investigate the possible mechanisms, P‑glycoprotein (P‑gp) protein expression was measured by western blotting and immunofluorescence assays. The mRNA expression levels of AKT, Erk1/2, cyclin‑dependent kinase inhibitor 1 (P21), caspase 3, cleaved caspase 3, caspase 9, cleaved caspase 9, poly (ADP‑ribose) polymerase (PARP), cleaved PARP, matrix metalloproteinase (MMP)‑2 and MMP‑9 were examined by reverse transcription‑quantitative (RT‑q) PCR assay, and the protein expression levels of phosphorylated (p)‑AKT, p‑Erk1/2, P21, caspase 3, cleaved caspase 3, caspase 9, cleaved caspase 9, PARP, cleaved PARP, MMP‑2 and MMP‑9 proteins expression by western blot assay. The MTS assay demonstrated that HNK (5 µg/ml), CUR (10 µg/ml) and DDP (5 µg/ml) had no obvious toxicity to A549/DDP cells, and HNK, CUR and DDP were more sensitive in A549 cells compared with A549/DDP cells. The optimal concentrations of HNK (5 µg/ml), CUR (10 µg/ml) and DDP (5 µg/ml) were chosen to carry out the further experiments. Compared with the control group, no significant change was observed in cell proliferation, apoptosis, migration, invasion and related mRNA and protein expression in HNK, CUR, DDP and HNK + CUR groups. The cell proliferation rate in the HNK + DDP and CUR + DDP groups was significantly suppressed with cell apoptosis significantly increased, respectively. The invasion cell number and wound healing rate of HNK + DDP and CUR + DDP groups were significantly depressed compared with the control group, respectively. Immunofluorescence demonstrated that the nuclear volume of P‑gp in HNK + DDP and CUR + DDP groups were significantly downregulated compared with the control group, respectively. The RT‑qPCR assay demonstrated that the AKT, Erk1/2 and P21 mRNA expression levels were significantly decreased and cleaved caspase 3, cleaved caspase 9 and cleaved PARP were increased in HNK + DDP and CUR + DDP groups compared with the control group. The western blotting results were consistent with the RT‑qPCR results. NK + CUR + DDP had improved effects on A549/DDP compared with HNK + DDP or CUR + DDP group, respectively. HNK and/or CUR could improve the sensitivity of DDP to A549/DDP cell by the regulation of P‑gp, inducing apoptosis, and inhibiting migration and invasion via AKT/ERK signal pathway in an in vitro study.

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1	Zheng R, Zeng H, Zhang S, Fan Y, Qiao Y, Zhou Q and Chen W: Lung cancer incidence and mortality in China, 2010. Thorac Cancer. 5:330–336. 2014.PubMed/NCBI View Article : Google Scholar
2	She J, Yang P, Hong Q and Bai C: Lung cancer in China: Challenges and interventions. Chest. 143:1117–1126. 2013.PubMed/NCBI View Article : Google Scholar
3	Schwartz AM and Rezaei MK: Diagnostic surgical pathology in lung cancer: Diagnosis and management of lung cancer, 3rd ed: American College of Chest Physicians evidence-based clinical practice guidelines. Chest. 143 (Suppl 5):Se251–Se262. 2013.PubMed/NCBI View Article : Google Scholar
4	Collins LG, Haines C, Perkel R and Enck RE: Lung cancer: Diagnosis and management. Am Fam Physician. 75:56–63. 2007.PubMed/NCBI
5	Goldstraw P, Ball D, Jett JR, Le Chevalier T, Lim E, Nicholson AG and Shepherd FA: Non-small-cell lung cancer. Lancet. 378:1727–1740. 2011.PubMed/NCBI View Article : Google Scholar
6	Fennell DA, Summers Y, Cadranel J, Benepal T, Christoph DC, Lal R, Das M, Maxwell F, Visseren-Grul C and Ferry D: Cisplatin in the modern era: The backbone of first-line chemotherapy for non-small cell lung cancer. Cancer Treat Rev. 44:42–50. 2016.PubMed/NCBI View Article : Google Scholar
7	Amable L: Cisplatin resistance and opportunities for precision medicine. Pharmacol Res. 106:27–36. 2016.PubMed/NCBI View Article : Google Scholar
8	Holford J, Sharp SY, Murrer BA, Abrams M and Kelland LR: In vitro circumvention of cisplatin resistance by the novel sterically hindered platinum complex AMD473. Br J Cancer. 77:366–373. 1998.PubMed/NCBI View Article : Google Scholar
9	Averett C, Arora S, Zubair H, Singh S, Bhardwaj A and Singh AP: Molecular targets of Honokiol: A promising phytochemical for effective cancer management. Enzymes. 36:175–193. 2014.PubMed/NCBI View Article : Google Scholar
10	Park E, Min H, Chung H, Hong J, Kang Y, Hung T, Youn U, Kim Y, Bae K, Kang S and Lee S: Down-regulation of c-Src/EGFR-mediated signaling activation is involved in the honokiol-induced cell cycle arrest and apoptosis in MDA-MB-231 human breast cancer cells. Cancer Lett. 277:133–140. 2009.PubMed/NCBI View Article : Google Scholar
11	Fan Y, Xue W, Schachner M and Zhao W: Honokiol eliminates glioma/glioblastoma stem cell-like cells via JAK-STAT3 signaling and inhibits tumor progression by targeting epidermal growth factor receptor. Cancers (Basel). 11(22)2018.PubMed/NCBI View Article : Google Scholar
12	Sengupta S, Nagalingam A, Muniraj N, Bonner MY, Mistriotis P, Afthinos A, Kuppusamy P, Lanoue D, Cho S, Korangath P, et al: Activation of tumor suppressor LKB1 by honokiol abrogates cancer stem-like phenotype in breast cancer via inhibition of oncogenic Stat3. Oncogene. 36:5709–5721. 2017.PubMed/NCBI View Article : Google Scholar
13	Wu H, Yin Z, Wang L, Li F and Qiu Y: Honokiol improved chondrogenesis and suppressed inflammation in human umbilical cord derived mesenchymal stem cells via blocking nuclear factor-κB pathway. BMC Cell Biol. 18(29)2017.PubMed/NCBI View Article : Google Scholar
14	Li Y and Zhang T: Targeting cancer stem cells by curcumin and clinical applications. Cancer Lett. 346:197–205. 2014.PubMed/NCBI View Article : Google Scholar
15	Cheng Q, Liao M, Hu H, Li H and Wu L: Asiatic acid (AA) sensitizes multidrug-resistant human lung adenocarcinoma A549/DDP cells to cisplatin (DDP) via downregulation of P-Glycoprotein (MDR1) and its targets. Cell Physiol Biochem. 47:279–292. 2018.PubMed/NCBI View Article : Google Scholar
16	Chen LP, Wang P, Sun YJ and Wu YJ: Direct interaction of avermectin with epidermal growth factor receptor mediates the penetration resistance in Drosophila larvae. Open Biol. 6(150231)2016.PubMed/NCBI View Article : Google Scholar
17	Liu Z, Sun M, Lu K, Liu J, Zhang M, Wu W, De W, Wang Z and Wang R: The long noncoding RNA HOTAIR contributes to cisplatin resistance of human lung adenocarcinoma cells via downregualtion of p21WAF1/CIP1 expression. PLoS One. 8:e77293–e77303. 2013.PubMed/NCBI View Article : Google Scholar
18	Lin Y, Duan Q and Yang YP: Immunohistochemistry of phosphatase and tensin homolog and metalloproteinase-9 in breast invasive micropapillary carcinoma. Eur J Gynaecol Oncol. 40:380–383. 2019.
19	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001.PubMed/NCBI View Article : Google Scholar
20	Dong Z, Ren L, Lin L and Li J, Huang Y and Li J: Effect of microRNA-21 on multidrug resistance reversal in A549/DDP human lung cancer cells. Mol Med Rep. 11:682–690. 2014.PubMed/NCBI View Article : Google Scholar
21	Xu D, Lu Q and Hu X: Down-regulation of P-glycoprotein expression in MDR breast cancer cell MCF-7/ADR by honokiol. Cancer Lett. 243:274–280. 2006.PubMed/NCBI View Article : Google Scholar
22	Ishitsuka K, Hideshima T, Hamasaki M, Raje N, Kumar S, Hideshima H, Shiraishi N, Yasui H, Roccaro AM, Richardson P, et al: Honokiol overcomes conventional drug resistance in human multiple myeloma by induction of caspase-dependent and -independent apoptosis. Blood. 106:1794–1800. 2005.PubMed/NCBI View Article : Google Scholar
23	Lee S, Khoo C, Halstead CW, Huynh T and Bensoussan A: Liquid chromatographic determination of honokiol and magnolol in hou po (Magnolia officinalis) as the raw herb and dried aqueous extract. J AOAC Int. 90:1210–1218. 2019.PubMed/NCBI
24	Chio CC, Chen KY, Chang CK, Chuang JY, Liu CC, Liu SH and Chen RM: Improved effects of honokiol on temozolomide-induced autophagy and apoptosis of drug-sensitive and -tolerant glioma cells. BMC Cancer. 18(379)2018.PubMed/NCBI View Article : Google Scholar
25	Li W, Wang Q, Su Q, Ma D, An C, Ma L and Liang H: Honokiol suppresses renal cancer cells' metastasis via dual-blocking epithelial-mesenchymal transition and cancer stem cell properties through modulating miR-141/ZEB2 signaling. Mol Cells. 37:383–388. 2014.PubMed/NCBI View Article : Google Scholar
26	Yao C, Lai G, Yeh C, Lai M, Shih P, Chao W, Whang-Peng J, Chuang S and Lai T: Honokiol eliminates human oral cancer stem-like cells accompanied with suppression of Wnt/β-catenin signaling and apoptosis induction. Evid Based Complement Alternat Med. 2013(146136)2013.PubMed/NCBI View Article : Google Scholar
27	Wynn M, Consul N, Merajver S and Schnell S: Inferring the effects of honokiol on the notch signaling pathway in SW480 colon cancer cells. Cancer Inform. 13 (Suppl 5):S1–S12. 2014.PubMed/NCBI View Article : Google Scholar
28	Lai I, Shih P, Yao C, Yeh C, Wang-Peng J, Lui T, Chuang S, Hu T, Lai T and Lai G: Elimination of cancer stem-like cells and potentiation of temozolomide sensitivity by Honokiol in glioblastoma multiforme cells. PLoS One. 10(e0114830)2015.PubMed/NCBI View Article : Google Scholar
29	Ye M and Zhang J and Zhang J, Miao Q, Yao L and Zhang J: Curcumin promotes apoptosis by activating the p53-miR-1922-5p/215-XIAP pathway in non-small cell lung cancer. Cancer Lett. 357:196–205. 2015.PubMed/NCBI View Article : Google Scholar
30	Lv L, Qiu K, Yu X, Chen C, Qin F, Shi Y, Ou J, Zhang T, Zhu H, Wu J, et al: Amphiphilic copolymeric micelles for doxorubicin and curcumin co-delivery to reverse multidrug resistance in breast cancer. J Biomed Nanotechnol. 12:973–985. 2016.PubMed/NCBI View Article : Google Scholar
31	Yuan HB, Meng PY and Qi LJ: Curcumin up-regulates miR-133a expression to inhibit hepatocellular carcinoma cell migration and invasion. World Chinese J Digestol, 2019.
32	Hosseini A, Rasmi Y, Rahbarghazi R, Aramwit P, Daeihassani B and Saboory E: Curcumin modulates the angiogenic potential of human endothelial cells via FAK/P-38 MAPK signaling pathway. Gene. 688:7–12. 2019.PubMed/NCBI View Article : Google Scholar
33	Almanaa T, Geusz M and Jamasbi R: Effects of curcumin on stem-like cells in human esophageal squamous carcinoma cell lines. BMC Complement Altern Med. 12(195)2012.PubMed/NCBI View Article : Google Scholar
34	Huang Y, Lin Y, Chiu H and Chiang B: Curcumin induces apoptosis of colorectal cancer stem cells by coupling with CD44 marker. J Agric Food Chem. 64:2247–2253. 2016.PubMed/NCBI View Article : Google Scholar
35	Mukherjee S, Mazumdar M, Chakraborty S, Manna A, Saha S, Khan P, Bhattacharjee P, Guha D, Adhikary A, Mukhjerjee S and Das T: Curcumin inhibits breast cancer stem cell migration by amplifying the E-cadherin/β-catenin negative feedback loop. Stem Cell Res Ther. 5(116)2014.PubMed/NCBI View Article : Google Scholar
36	Wilkens S: Structure and mechanism of ABC transporters. F1000Prime Rep. 7(14)2015.PubMed/NCBI View Article : Google Scholar
37	Liu CH, Huang Q, Jin ZY, Zhu CL, Liu Z and Wang C: miR-21 and KLF4 jointly augment epithelial-mesenchymal transition via the Akt/ERK1/2 pathway. Int J Oncol. 50:1109–1115. 2017.PubMed/NCBI View Article : Google Scholar
38	Zhang S, Bian H, Li X, Wu H, Bi Q, Yan Y and Wang Y: Hydrogen sulfide promotes cell proliferation of oral cancer through activation of the COX2/AKT/ERK1/2 axis. Oncol Rep. 35:2825–2832. 2016.PubMed/NCBI View Article : Google Scholar
39	Krebs K, Ruusmann A, Simonlatser G and Velling T: Expression of FLNa in human melanoma cells regulates the function of integrin α1β1 and phosphorylation and localisation of PKB/AKT/ERK1/2 kinases. Eur J Cell Biol. 94:564–575. 2015.PubMed/NCBI View Article : Google Scholar
40	Hu Y, Hong Y, Xu Y, Liu P, Guo DH and Chen Y: Inhibition of the JAK/STAT pathway with ruxolitinib overcomes cisplatin resistance in non-small-cell lung cancer NSCLC. Apoptosis. 19:1627–1636. 2014.PubMed/NCBI View Article : Google Scholar
41	Zhang W, Zhou H, Yu Y, Li J, Li H, Jiang D, Chen Z, Yang D, Xu Z and Yu Z: Combination of gambogic acid with cisplatin enhances the antitumor effects on cisplatin-resistant lung cancer cells by downregulating MRP2 and LRP expression. Onco Targets Ther. 9:3359–3368. 2016.PubMed/NCBI View Article : Google Scholar
42	Dai M, Alodaini AA, Filsaimé N, Villatoro MA, Guo J, Arakelian A, Rabbani SA, Ali S and Lebrun JJ: Cyclin D1 cooperates with p21 to regulate TGFβ-mediated breast cancer cell migration and tumor local invasion. Breast Cancer Res. 15(3246)2013.PubMed/NCBI View Article : Google Scholar
43	Giganti Μg, Tresoldi I, Sorge R, Melchiorri G, Triossi T, Masuelli L, Lido P, Albonici L, Foti C, Modesti A and Bei R: Physical exercise modulates the level of serum MMP-2 and MMP-9 in patients with breast cancer. Oncol Lett. 12:2119–2126. 2016.PubMed/NCBI View Article : Google Scholar
44	Jayakumar T, Liu CH, Wu GY, Lee TY, Manubolu M, Hsieh CY, Yang CH and Sheu JR: Hinokitiol inhibits migration of A549 lung cancer cells via Suppression of MMPs and induction of antioxidant enzymes and apoptosis. Int J Mol Sci. 19(939)2018.PubMed/NCBI View Article : Google Scholar
45	Laskowska M: Altered maternal serum matrix metalloproteinases MMP-2, MMP-3, MMP-9, and MMP-13 in severe early- and late-onset preeclampsia. Biomed Res Int. 2017(6432426)2017.PubMed/NCBI View Article : Google Scholar
46	Chen J, Ren Z, Zhu M and Khalil RA: Decreased homodimerization and increased TIMP-1 complexation of uteroplacental and uterine arterial matrix metalloproteinase-9 during hypertension-in-pregnancy. Biochem Pharmacol. 138:81–95. 2017.PubMed/NCBI View Article : Google Scholar