Abstract
Objective
To investigate the cellular effects of Pien Tze Huang (片仔癀, PZH) in the HT-29 human colon carcinoma cell line.
Methods
The viability of HT-29 cells was determined by MTT assay. A fluorescence-activated cell sorting (FACS) analysis with annexin-V/propidium iodide (PI) and JC-1 staining were performed to determine cell apoptosis and the loss of mitochondrial membrane potential, respectively. Activation of caspase 3 was evaluated by a colorimetric assay. The mRNA expression levels of Bcl-2 and Bax were measured by reverse transcription polymerase chain reaction (RT-PCR).
Results
PZH, in a dose- and timedependent manner, reduced viability and induced apoptosis of HT-29 cells. Moreover, PZH treatment resulted in the collapse of the mitochondrial membrane potential, activation of caspase 3, and an increase in the Bax/Bcl-2 ratio.
Conclusion
PZH inhibits the growth of HT-29 cells by inducing cancer cell apoptosis via regulation of the Bcl-2 family and activation of caspase 3, which may, in part, explain its anticancer activity.
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References
Adams JM, Cory S. The Bcl-2 apoptotic switch in cancer development and therapy. Oncogene 2007;26:1324–1337.
Cory S, Adams JM. The Bcl-2 family: regulators of the cellular life-of-death switch. Nat Rev Cancer 2002;2:647–656.
Hsu YT, Wolter K, Youle RJ. Cytosol to membrane redistribution of members of the Bcl-2 family during apoptosis. Proc Natl Acad Sci USA 1997;94:3668–3672.
Wolter KG, Hsu YT, Smith CL, Nechushtan A, Xi XG, Youle RJ. Movement of Bax from the cytosol to mitochondria. J Cell Biol 1997;139:1281–1292.
Kluck RM, Bossy-Wetzel E, Green DR, Newmeyer DD. The release of cytochrome C from mitochondria: a primary site for Bcl-2 regulation of apoptosis. Science 1997;275:1132–1136.
Jürgensmeier JM, Xie Z, Deveraux Q, Ellerby L, Bredesen D, Reed JC. Bax directly induces release of cytochrome C from isolated mitochondria. Proc Natl Acad Sci USA 1998;95:4997–5002.
Antonsson B, Montessuit S, Lauper S, Eskes R, Martinou JC. Bax oligomerization is required for channel-forming activity in liposomes and to trigger cytochrome C release from mitochondria. Biochem J 2000;345:271–278.
Gross A, McDonnell JM, Korsmeyer SJ. Bcl-2 family members and the mitochondria in apoptosis. Genes Dev 1999;13:1899–1911.
Yang J, Liu X, Bhalla K, Kim CN, Ibrado AM, Cai J, et al. Prevention of apoptosis by Bcl-2: release of cytochrome C from mitochondria blocked. Science 1997;275:1129–1132.
Thomenius MJ, Wang NS, Reineks EZ, Wang Z, Distelhorst CW. Bcl-2 on the endoplasmic reticulum regulates Bax activity by binding to BH3-only proteins. J Biol Chem 2003;278:6243–6250.
Antonsson B, Conti F, Ciavatta A, Montessuit S, Lewis S, Martinou I, et al. Inhibition of Bax channel-forming activity by Bcl-2. Science 1997;277:370–372.
Peng J, Ding J, Tan C, Baggenstoss B, Zhang Z, Lapolla SM, et al. Oligomerization of membrane-bound Bcl-2 is involved in its pore formation induced by tBid. Apoptosis 2009;14:1145–1153.
Peng J, Tan C, Roberts GJ, Nikolaeva O, Lapolla SM, Andrews DW, et al. tBid elicits a conformational alteration in membrane-bound Bcl-2 such that it inhibits Bax pore formation. J Biol Chem 2006;281:35802–35811.
Youle RJ, Strasser A. The Bcl-2 protein family: opposing activities that mediate cell death. Nat Rev Mol Cell Biol 2008;9:47–59.
Yip KW, Reed JC. Bcl-2 family proteins and cancer. Oncogene 2008;27:6398–6406.
Chinese Pharmacopoeia Commission. Pharmacopoeia of the People's Republic of China. Beijing: China Medical Science and Technology Press; 2010:573–575.
Lee KK, Kwong WH, Chau FT, Yew DT, Chan WY. Pien Tze Huang protects the liver against carbon tetrachloride induced damage. Pharmacol Toxicol 2002;91:185–192.
Xu YY, Yu EX. Clinical analysis of the effect of Pien Tze Huang in treatment of 42 patients with moderate or advanced liver cancer. Shanghai J Tradit Chin Med (Chin) 1994;12:4–5.
Gu ZX. Therapeutical observation of advanced colon cancer. Chin Tradit Patent Med (Chin) 1993;15; 23.
Liu CS. Review of pharmacology and clinical application of Pien Tze Huang. Med Pharm World 2006;7:64–66.
Lin JM, Chen YQ, Wei LH, Chen XZ, Xu W, Hong ZF, et al. Hedyotis Diffusa Willd extract induces apoptosis via activation of the mitochondrion-dependent pathway in human colon carcinoma cells. Int J Oncol 2010;37:1331–1338.
Mantymaa P, Siitonen T, Guttorm T, Saily M, Kinnula V, Savolainen ER, et al. Induction of mitochondrial manganese superoxide dismutase confers resistance to apoptosis in acute myeloblastic leukaemia cells exposed to etoposide. Br J Haematol 2000;108:574–581.
Korper S, Nolte F, Rojewski MT, Thiel E, Schrezenmeier H. The K+ channel openers diazoxide and NS1619 induce depolarization of mitochondria and have differential effects on cell Ca2+ in CD34+ cell line KG-1. Exp Hematol 2003;31:815–823.
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Supported by the National Natural Science Foundation of China (No. 81073097), the Developmental Fund of CHEN Ke-ji Integrative Medicine (No. CKJ 2011001), and the Natural Science Foundation of Fujian Province of China (No. 2010J01195)
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Lin, Jm., Wei, Lh., Chen, Yq. et al. Pien Tze Huang (片仔癀)-induced apoptosis in human colon cancer HT-29 cells is associated with regulation of the Bcl-2 family and activation of caspase 3. Chin. J. Integr. Med. 17, 685–690 (2011). https://doi.org/10.1007/s11655-011-0846-4
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DOI: https://doi.org/10.1007/s11655-011-0846-4