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Bauhinia variegata Bark Extract: Assessment of its Anti-proliferative and Apoptotic Activities on A549 and H460 Lung Cancer Cell Lines


Affiliations
1 Department of Biochemistry, The Maharaja Sayajirao University of Baroda, Vadodara - 390002, Gujarat, India
 

The hunt for novel anticancer drugs with minimal side effects continues. This study strengthens the claim by providing biochemical evidences of anticancer activities of Bauhinia variegata bark extracts on lung carcinoma cells (A549 and H460). Bark extracts of Bauhinia variegata were prepared by different solvents using Soxhlet apparatus and tested for their antioxidant potential by DPPH assay. The lung cancer cell lines were treated with Bauhinia variegata bark extracts and viability of cells was measured by MTT assay; metastatic ability was determined through Scratch assay and effect on DNA integrity was shown by gel electrophoresis. The Petroleum Ether Bark Extract (PEBE) inhibits proliferation (A549, IC50 = 1.5 mg/ml) at 48 h treatment. DNA damage was observed in A549 cells by agarose gel electrophoresis. The Chloroform Bark Extract (CBE) inhibited proliferation of H460 (IC50 = 1 mg/ml) with DNA damage after 24 h treatment. Soft agar assay indicated decreased ability to form colonies and scratch test showed impaired migration of A549 and H460 to PEBE and CBE treatment respectively. Apoptosis was detected using fluorescent dye staining in A549 and H460 cells. Caspase 3 activity was increased significantly in A549 and H460 cells. PEBE and CBE decrease the mitochondrial membrane potential gradient (&#916&#936m) of A549 and H460 cells respectively. This study categorically proves the cytotoxic activity of Bauhinia variegata bark extracts on A549 and H460 cells.

Keywords

Anticancer Effect, Caspase, DNA Damage, Metastasis, Non-small Cell Lung Cancer Cell Lines
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  • Parikh PM, Ranade AA, Govind B, Ghadyalpatil N, Singh R, Bharath R, et al. Lung cancer in India: Current status and promising strategies. South Asian J Cancer. 2016; 5(03):093-5. https://doi.org/10.4103/2278-330X.187563. PMid:27606289. PMCid:PMC4991145
  • Zappa C, Mousa SA. Non-small cell lung cancer: Current treatment and future advances. Transl Lung Cancer Res. 2016; 5(3):288. https://doi.org/10.21037/tlcr.2016.06.07. PMid:27413711. PMCid:PMC4931124
  • Lu T, Yang X, Huang Y, Zhao M, Li M, Ma K, et al. Trends in the incidence, treatment, and survival of patients with lung cancer in the last four decades. Cancer Manag Res. 2019; 11:943. https://doi.org/10.2147/CMAR.S187317. PMid:30718965. PMCid:PMC6345192
  • Debevec L, Debeljak A. Multidisciplinary management of lung cancer. J Thorac Oncol. 2007; 2(6):577. https://doi.org/10.1097/JTO.0b013e318060f16d. PMid:17545858
  • Huang CY, Ju DT, Chang CF, Reddy PM, Velmurugan BK. A review on the effects of current chemotherapy drugs and natural agents in treating non-small cell lung cancer. Biomedicine. 2017; 7(4). https://doi.org/10.1051/bmdcn/2017070423. PMid:29130448. PMCid:PMC5682982
  • Jin S, Park HJ, Oh YN, Kwon HJ, Kim JH, Choi YH, et al. Anti-cancer activity of Osmanthus matsumuranus extract by inducing G2/M arrest and apoptosis in human hepatocellular carcinoma Hep G2 cells. J Cancer Prev. 2015; 20(4):241. https://doi.org/10.15430/JCP.2015.20.4.241.
  • PMid:26734586. PMCid:PMC4699751
  • Richardson JS, Sethi G, Lee GS, Malek SN. Chalepin: isolated from Ruta angustifolia L. Pers induces mitochondrial mediated apoptosis in lung carcinoma cells. BMC Complement Altern Med. 2016; 16(1):1-27. https://doi.org/10.1186/s12906-016-1368-6. PMid:27729078
  • PMCid:PMC5059921
  • Mali RG, Dhake AS. Evaluation of effects of Bauhinia variegata stem bark extracts against milk-induced eosinophilia in mice. J. Adv. Pharm. Technol. Res. 2011; 2(2):132. https://doi.org/10.4103/2231-4040.82949. PMid:22171306. PMCid:PMC3217693
  • Katoch D, Sharma JS, Banerjee S, Biswas R, Das B, Goswami D, et al. Government policies and initiatives for development of Ayurveda. J Ethnopharmacol. 2017; 197:25-31. https://doi.org/10.1016/j.jep.2016.08.018. PMid:27543425
  • Aggarwal V, Tuli HS, Varol A, Thakral F, Yerer MB, Sak K, et al. Role of reactive oxygen species in cancer progression: Molecular mechanisms and recent advancements. Biomolecules. 2019; 9(11):735. https://doi.org/10.3390/biom9110735. PMid:31766246. PMCid:PMC6920770
  • Galadari S, Rahman A, Pallichankandy S, Thayyullathil F. Reactive oxygen species and cancer paradox: To promote or to suppress? Free Radic Biol Med. 2017; 104:144-64. https://doi.org/10.1016/j.freeradbiomed.2017.01.004.
  • PMid:28088622
  • Kirtonia A, Sethi G, Garg M. The multifaceted role of reactive oxygen species in tumorigenesis. Cell Mol Life Sci. 2020:1-25.
  • Hensley P, Mishra M, Kyprianou N. Targeting caspases in cancer therapeutics. Biol. Chem. 2013; 394(7):831-43. https://doi.org/10.1515/hsz-2013-0128. PMid:23509217. PMCid:PMC3721733
  • Jelínek M, Balušíková K, Schmiedlová M, NěmcováFürstová V, Šrámek J, Stančíková J, et al. The role of individual caspases in cell death induction by taxanes in breast cancer cells. Cancer Cell Int. 2015; 15(1):1-6. https://doi.org/10.1186/s12935-015-0155-7. PMid:25685064.
  • PMCid:PMC4329194
  • Olsson M, Zhivotovsky B. Caspases and cancer. Cell Death Differ. 2011; 18(9):1441-9. https://doi.org/10.1038/cdd.2011.30. PMid:21455218. PMCid:PMC3178435
  • Khazaei M, Pazhouhi M. Antiproliferative effect of Trifolium pratens L. extract in human breast cancer cells. Nutr Cancer.2019; 71(1):128-40. https://doi.org/10.1080/01635581.2018.1521443. PMid:30596276
  • Bhandari J, Muhammad B, Thapa P, Shrestha BG. Study of phytochemical, anti-microbial, anti-oxidant, and anti-cancer properties of Allium wallichii. BMC Complement Altern Med. 2017; 17(1):1-9. https://doi.org/10.1186/s12906-017-1622-6. PMid:28178952. PMCid:PMC5299666
  • Kothari S, Mishra V, Bharat S, Tonpay SD. Antimicrobial activity and phytochemical screening of serial extracts from leaves of Aegle marmelos (Linn.). Acta Pol. Pharm. 2011;68(5):687-92.
  • Zhao T, Pan H, Feng Y, Li H, Zhao Y. Petroleum ether extract of Chenopodium album L. prevents cell growth and induces apoptosis of human lung cancer cells. Exp Ther Med. 2016;12(5):3301-7. https://doi.org/10.3892/etm.2016.3765. PMid:27882153. PMCid:PMC5103781
  • Franken NA, Rodermond HM, Stap J, Haveman J, Van Bree C. Clonogenic assay of cells in vitro. Nat Protoc. 2006; 1(5):2315-9. https://doi.org/10.1038/nprot.2006.339. PMid:17406473
  • Liang CC, Park AY, Guan JL. In vitro scratch assay: A convenient and inexpensive method for analysis of cell migration in vitro. Nat Protoc. 2007; 2(2):329-33. https://doi.org/10.1038/nprot.2007.30. PMid:17406593
  • Liu N, Li Y, Su S, Wang N, Wang H, Li J. Inhibition of cell migration by ouabain in the A549 human lung cancer cell line. Oncol Lett. 2013; 6(2):475-9. https://doi.org/10.3892/ol.2013.1406. PMid:24137350. PMCid:PMC3789103
  • Hsia TC, Yu CC, Hsu SC, Tang NY, Lu HF, Huang YP, et al. Cantharidin induces apoptosis of H460 human lung cancer cells through mitochondria-dependent pathways. Int J Oncol. 2014; 45(1):245-54. https://doi.org/10.3892/ijo.2014.2428. PMid:24818581
  • Lin W, Ye H. Anticancer activity of ursolic acid on human ovarian cancer cells via ROS and MMP mediated apoptosis, cell cycle arrest and downregulation of PI3K/AKT pathway. J Buon. 2020; 25(2):750-56.
  • Alizadeh J, Zeki AA, Mirzaei N, Tewary S, Moghadam AR, et al. Mevalonate cascade inhibition by simvastatin induces the intrinsic apoptosis pathway via depletion of isoprenoids in tumor cells. Sci Rep. 2017; 7(1):1-4. https://doi.org/10.1038/srep44841. PMid:28344327. PMCid:PMC5366866
  • Bang M, Do Gyeong Kim EL, Kwon KJ, Shin CY. Etoposide induces mitochondrial dysfunction and cellular senescence in primary cultured rat astrocytes. Biomol Ther. 2019;27(6):530. https://doi.org/10.4062/biomolther.2019.151. PMid:31646843. PMCid:PMC6824621
  • Elmore S. Apoptosis: A review of programmed cell death. Toxicol Pathol. 2007; 35(4):495-516.
  • https://doi. org/10.1080/01926230701320337. PMid:17562483. PMCid:PMC2117903
  • Perillo B, Di Donato M, Pezone A, Di Zazzo E, Giovannelli P, Galasso G, et al. ROS in cancer therapy: The bright side of the moon. Exp Mol Med. 2020; 52(2):192-203. https://doi.org/10.1038/s12276-020-0384-2. PMid:32060354 PMCid:PMC7062874
  • Clark AC, MacKenzie SH. Targeting cell death in tumors by activating caspases. Curr Cancer Drug Targets. 2008;8(2):98-109. https://doi.org/10.2174/156800908783769391 PMid:18336192. PMCid:PMC3119715
  • Kumar S. Caspase function in programmed cell death. Cell Death Differ. 2007; 14(1):32-43. https://doi.org/10.1038/sj.cdd.4402060. PMid:17082813
  • Ji BC, Yu CC, Yang ST, Hsia TC, Yang JS, Lai KC, et al. Induction of DNA damage by deguelin is mediated through reducing DNA repair genes in human non-small cell lung cancer NCI-H460 cells. Oncol. Rep. 2012; 27(4):959-64. https://doi.org/10.3892/or.2012.1622. PMid:22227970.
  • PMCid:PMC3583480
  • Tiloke C, Phulukdaree A, Chuturgoon AA. The antiproliferative effect of Moringa oleifera crude aqueous leaf extract on cancerous human alveolar epithelial cells. BMC Complement Altern Med. 2013; 13(1):1-8. https://
  • doi.org/10.1186/1472-6882-13-226. PMid:24041017. PMCid:PMC3852616

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  • Bauhinia variegata Bark Extract: Assessment of its Anti-proliferative and Apoptotic Activities on A549 and H460 Lung Cancer Cell Lines

Abstract Views: 24  |  PDF Views: 7

Authors

Tanvi Khanna
Department of Biochemistry, The Maharaja Sayajirao University of Baroda, Vadodara - 390002, Gujarat, India
Akash Dave
Department of Biochemistry, The Maharaja Sayajirao University of Baroda, Vadodara - 390002, Gujarat, India
Sejal Purani
Department of Biochemistry, The Maharaja Sayajirao University of Baroda, Vadodara - 390002, Gujarat, India
Jagath Vedamurthy
Department of Biochemistry, The Maharaja Sayajirao University of Baroda, Vadodara - 390002, Gujarat, India
Dhaval Jivani
Department of Biochemistry, The Maharaja Sayajirao University of Baroda, Vadodara - 390002, Gujarat, India
Pushpa Robin
Department of Biochemistry, The Maharaja Sayajirao University of Baroda, Vadodara - 390002, Gujarat, India

Abstract


The hunt for novel anticancer drugs with minimal side effects continues. This study strengthens the claim by providing biochemical evidences of anticancer activities of Bauhinia variegata bark extracts on lung carcinoma cells (A549 and H460). Bark extracts of Bauhinia variegata were prepared by different solvents using Soxhlet apparatus and tested for their antioxidant potential by DPPH assay. The lung cancer cell lines were treated with Bauhinia variegata bark extracts and viability of cells was measured by MTT assay; metastatic ability was determined through Scratch assay and effect on DNA integrity was shown by gel electrophoresis. The Petroleum Ether Bark Extract (PEBE) inhibits proliferation (A549, IC50 = 1.5 mg/ml) at 48 h treatment. DNA damage was observed in A549 cells by agarose gel electrophoresis. The Chloroform Bark Extract (CBE) inhibited proliferation of H460 (IC50 = 1 mg/ml) with DNA damage after 24 h treatment. Soft agar assay indicated decreased ability to form colonies and scratch test showed impaired migration of A549 and H460 to PEBE and CBE treatment respectively. Apoptosis was detected using fluorescent dye staining in A549 and H460 cells. Caspase 3 activity was increased significantly in A549 and H460 cells. PEBE and CBE decrease the mitochondrial membrane potential gradient (&#916&#936m) of A549 and H460 cells respectively. This study categorically proves the cytotoxic activity of Bauhinia variegata bark extracts on A549 and H460 cells.

Keywords


Anticancer Effect, Caspase, DNA Damage, Metastasis, Non-small Cell Lung Cancer Cell Lines

References





DOI: https://doi.org/10.18311/jnr%2F2022%2F28740