Browse > Article
http://dx.doi.org/10.7314/APJCP.2013.14.2.631

Comparative Studies to Evaluate Relative in vitro Potency of Luteolin in Inducing Cell Cycle Arrest and Apoptosis in HaCaT and A375 Cells  

George, Vazhapilly Cijo (Cell Culture Laboratory, School of BioSciences and Technology, VIT University)
Kumar, Devanga Ragupathi Naveen (Cell Culture Laboratory, School of BioSciences and Technology, VIT University)
Suresh, Palamadai Krishnan (Cell Culture Laboratory, School of BioSciences and Technology, VIT University)
Kumar, Sanjay (Center for Stem Cell Research, Christian Medical College)
Kumar, Rangasamy Ashok (Department of Zoology, Government Arts College)
Publication Information
Asian Pacific Journal of Cancer Prevention / v.14, no.2, 2013 , pp. 631-637 More about this Journal
Abstract
Luteolin is a naturally occurring flavonoid present in many plants with diverse applications in pharmacology. Despite several studies elucidating its significant anti-cancer activity against various cancer cells, the mechanism of action in skin cancer is not well addressed. Hence, we investigated the effects of luteolin in HaCaT (human immortalized keratinocytes) and A375 (human melanoma) cells. The radical scavenging abilities of luteolin were determined spectrophotometrically, prior to a cytotoxic study (XTT assay). Inhibitory effects were assessed by colony formation assay. Further, the capability of luteolin to induce cell cycle arrest and apoptosis were demonstrated by flow cytometry and cellular DNA fragmentation ELISA, respectively. The results revealed that luteolin possesses considerable cytotoxicity against both HaCaT and A375 cells with $IC_{50}$ values of 37.1 ${\mu}M$ and 115.1 ${\mu}M$, respectively. Luteolin also inhibited colony formation and induced apoptosis in a dose and time-dependent manner by disturbing cellular integrity as evident from morphological evaluation by Wright-Giemsa staining. Accumulation of cells in G2/M (0.83-8.14%) phase for HaCaT cells and G0/G1 (60.4-72.6%) phase for A375 cells after 24 h treatment indicated cell cycle arresting potential of this flavonoid. These data suggest that luteolin inhibits cell proliferation and promotes cell cycle arrest and apoptosis in skin cancer cells with possible involvement of programmed cell death, providing a substantial basis for it to be developed into a potent chemopreventive template for skin cancer.
Keywords
Skin cancer; luteolin; cytotoxicity; apoptosis; cell cycle;
Citations & Related Records
Times Cited By KSCI : 2  (Citation Analysis)
연도 인용수 순위
1 Dua P, Gude RP (2006). Antiproliferative and antiproteolytic activity of pentoxifylline in cultures of B16F10 melanoma cells. Cancer Chemother Pharmacol, 58, 195-202.   DOI
2 Fabbrocini G, Triassi M, Mauriello MC, et al (2010). Epidemiology of skin cancer: role of some environmental factors. Cancers, 2, 1980-9.   DOI   ScienceOn
3 Fecker LF, Geilen CC, Hossini AM, et al (2005). Selective induction of apoptosis in melanoma cells by tyrosinase promoter-controlled CD95 ligand overexpression. J Invest Dermatol, 124, 221-8.   DOI   ScienceOn
4 Garbe C, Eigentler TK, Keilholz U, et al (2011). Systematic review of medical treatment in melanoma: current status and future prospects. The Oncologist, 16, 5-24.   DOI   ScienceOn
5 George VC, Kumar DRN, Rajkumar V, et al (2012). Quantitative assessment of the relative antineoplastic potential of the n-butanolic leaf extract of Annona muricata Linn. in normal and immortalized human cell lines. Asian Pac J Cancer Prev, 13, 699-704.   DOI   ScienceOn
6 Gibellini L, Pinti M, Nasi M, et al (2010). Interfering with ROS metabolism in cancer cells: the potential role of quercetin. Cancers, 2, 1288-311.   DOI   ScienceOn
7 Girschik J, Fritschi L, Threlfall T, et al (2008). Deaths from non-melanoma skin cancer in eastern Australia. Cancer Causes Control, 19, 879-85.   DOI
8 Green DR, Reed JC (1998). Mitochondria and apoptosis. Science, 281, 1309-12.   DOI   ScienceOn
9 Gu YH, Belury MA (2005). Selective induction of apoptosis in murine skin carcinoma cells (CH72) by an ethanol extract of Lentinula edodes. Cancer Lett, 220, 21-8.   DOI   ScienceOn
10 Hong WK, Sporn MB (1997). Recent advances in chemoprevention of cancer. Science, 278, 1073-7.   DOI   ScienceOn
11 Hsan KM, Chen CC, Shyur LF (2010). Current research and development of chemotherapeutic agents for melanoma. Cancers, 2, 397-419.   DOI   ScienceOn
12 Kajstura M, Halicka HD, Pryjma J, et al (2007). Discontinuous fragmentation of nuclear DNA during apoptosis revealed by discrete "sub-G1" peaks on DNA content histograms. Cytometry, 71, 125-31.
13 Kamran MZ, Gude RP (2012). Preclinical evaluation of the antimetastatic efficacy of Pentoxifylline on A375 human melanoma cell line. Biomed Pharmacother, 10, 6.
14 Kang N, Zhang JH, Qiu F, et al (2010). Induction of G(2)/M phase arrest and apoptosis by oridonin in human laryngeal carcinoma cells. J Nat Prod, 73, 1058-63.   DOI   ScienceOn
15 Kang OH, Choi JG, Lee JH, et al (2010). Luteolin isolated from the flowers of Lonicera japonica suppresses inflammatory mediator release by blocking NF-${\kappa}B$ and MAPKs activation pathways in HMC-1 cells. Molecules, 15, 385-98.   DOI   ScienceOn
16 Kim MJ, Woo JS, Kwon CH, et al (2012). Luteolin induces apoptotic cell death through AIF nuclear translocation mediated by activation of ERK and p38 in human breast cancer cell lines. Cell Biol Int, 36, 339-44.   DOI   ScienceOn
17 Kobayashi T, Nakata T, Kuzumaki T (2002). Effect of flavonoids on cell cycle progression in prostate cancer cells. Cancer Lett, 176, 17-23.   DOI   ScienceOn
18 Krishnamurthy R, Maly DJ (2010). Biochemical mechanisms of resistance to small-molecule protein kinase inhibitors. ACS Chem Biol, 5, 121-38.   DOI   ScienceOn
19 Kuo JH, Chu YL, Yang JS, et al (2010). Cantharidin induces apoptosis in human bladder cancer TSGH 8301 cells through mitochondria-dependent signal pathways. Int J Oncol, 37, 1243-50.
20 Lo C, Lai TY, Yang JS, et al (2011). Gallic acid inhibits the migration and invasion of A375.S2 human melanoma cells through the inhibition of matrix metalloproteinase-2 and Ras. Melanoma Res, 21, 267-73.   DOI   ScienceOn
21 Maioli E, Torricelli C, Fortino V, et al (2009). Critical appraisal of the MTT assay in the presence of rottlerin and uncouplers. Biol Proced Online, 11, 227-40.   DOI   ScienceOn
22 Middleton EJ, Kandaswami C, Theoharides TC (2000). Effects of plant flavonoids on mammalian cells:implications for inflammation, heart disease and cancer. Pharmacol Rev, 52, 673-751.
23 Miimohammadsadegh A, Hassan M, Bardenheuer W, et al (2006). STAT5 phosphorylation in malignant melanoma is important for survival and is mediated through SRC and JAK1 kinases. J Invest Dermatol, 126, 2272-80.   DOI   ScienceOn
24 Miimohammadsadegh A, Mota R, Gustrau A, et al (2007). ERK1/2 is highly phosphorylated in melanoma metastases and protects melanoma cells from cisplatin-mediated apoptosis. J Invest Dermatol, 127, 2207-15.   DOI   ScienceOn
25 Ozgen U, Mavi A, Terzi Z, et al (2011). Relationship between chemical structure and antioxidant activity of luteolin and its glycosides isolated from Thymus. Rec Nat Prod, 5, 12-21.
26 Pelicano H, Carney D, Huang P (2004). ROS stress in cancer cells and therapeutic implications. Drug Resist Updat, 7, 97-110.   DOI   ScienceOn
27 Ross JA, Kasum CM (2002). Dietary flavonoids: bioavailability, metabolic effects, and safety. Annu Rev Nutr, 22, 19-34.   DOI   ScienceOn
28 Seelinger G, Merfort I, Schempp CM (2008). Anti-oxidant, anti-inflammatory and anti-allergic activities of luteolin. Planta Med, 74, 1667-77.   DOI   ScienceOn
29 Shahidi F (1997). Natural antioxidants: An overview. In natural antioxidants, chemistry, health effects and applications. Edited by Shahidi F. Champaign, AOCS Press: 1-11.
30 Shi J, Wei L (2007). Rho kinase in the regulation of cell death and survival. Arch Immunol Ther Exp, 55, 61-75.   DOI
31 Shi RX, Ong CN, Shen HM (2004). Luteolin sensitizes tumor necrosis factor-alpha-induced apoptosis in human tumor cells. Oncogene, 23, 7712-21.   DOI   ScienceOn
32 Shimada K, Fujikawa K, Yahara K, et al (1992). Antioxidative properties of xanthin and autooxidation of soybean oil in cyclodextrin emulsion. J Agric Food Chem, 40, 945-8.   DOI
33 Soares JR, Dins TCP, Cunha AP, et al (1997). Antioxidant activity of some extracts of Thymus zygis. Free Radic Res, 26, 469-78.   DOI   ScienceOn
34 Ueda H, Yamazaki C, Yamazaki M (2002). Luteolin as an anti-inflammatory and anti-allergic constituent of Perilla frutescens. Biol Pharm Bull, 25, 1197-202.   DOI   ScienceOn
35 Voland C, Bord A, Peleraux A, et al (2006). Repression of cell cycle-related proteins by oxaliplatin but not cisplatin in human colon cancer cells. Mol Cancer Ther, 5, 2149-57.   DOI   ScienceOn
36 Wang HY, Quan K, Jiang YL, et al (2010). Effect of Luteolin and its combination with chemotherapeutic drugs on cytotoxicity of cancer cells. J Zhejiang Uni Medi Sci, 39, 30-6.
37 Wang LM, Xie KP, Huo HN, et al (2012). Luteolin inhibits proliferation induced by IGF-1 pathway dependent ER$\alpha$ in human breast cancer MCF-7 cells. Asian Pac J Cancer Prev, 13, 1431-7.   DOI   ScienceOn
38 Wang Z, Tang X, Zhang Y, et al (2012). Lobaplatin induces apoptosis and arrests cell cycle progression in a human cholangio carcinoma cell line RBE. Biomed Pharmacother, 66, 161-6.   DOI   ScienceOn
39 Weislow OS, Kiser R, Fine DL, et al (1989). New soluble-formazan assay for HIV-1 cytopathic effects: application to high-flux screening of synthetic and natural products for AIDS-antiviral activity. J Natl Cancer Inst, 81, 577-86.   DOI
40 Xu K, Liu B, Ma Y, et al (2009). Physicochemical properties and antioxidant activities of luteolin-phospholipid complex. Molecules, 14, 3486-93.   DOI   ScienceOn
41 Chahar KM, Sharma N, Dobhal MP, et al (2011). Flavonoids: a versatile source of anticancer drugs. Pharmacognosy Rev, 5, 1-12.   DOI   ScienceOn
42 Adinarayan M, Krishnamurthy SP (2011). Clinicopathological evaluation of nonmelanoma skin cancer. Indian J Dermatol, 56, 670-2.   DOI   ScienceOn
43 Benzie IF, Strain JJ (1996). The ferric reducing ability of plasma (FRAP) as a measure of "antioxidant power": the FRAP assay. Anal Biochem, 239, 70-6.   DOI   ScienceOn
44 Brady MS, Kaushal A, KO C, et al (2011). Melanoma and other skin cancers. Cancer Management, 14, 1-32.
45 Chang J, Hsu Y, Kuo P, et al (2005). Increase of Bax/ Bcl-XL ratio and arrest of cell cycle by luteolin in immortalized human hepatoma cell line. Life Sci, 76, 1883-93.   DOI   ScienceOn
46 Chen T, Wong YS (2009). Selenocystine induces reactive oxygen species mediated apoptosis in human cancer cells. Biomed Pharmacother, 63, 105-13.   DOI   ScienceOn
47 Chiang CT, Way TD, Lin JK (2007). Sensitizing HER2-overexpressing cancer cells to luteolin-induced apoptosis through suppressing p21(WAF1/CIP1) expression with rapamycin. Mol Cancer Ther, 6, 2127-38.   DOI   ScienceOn
48 Chung H, Suh EK, Han IO, et al (2011). Keratinocyte-derived laminin-332 promotes adhesion and migration in melanocytes and melanoma. J Biol Chem, 15, 13438-47.
49 Deorukhkar A, Krishnan S, Sethi G (2007). Back to basics: how natural products can provide the basis for new therapeutics. Expert Opin Investig Drugs, 16, 1753-73.   DOI   ScienceOn