Caspase and mitogen activated protein kinase pathways are involved in Solanum lyratum herba induced apoptosis.
OBJECTIVE
Solanum lyratum herba (SLH) has been traditionally used for the treatment of febrifuge, diarrhea, eye disease and cancer with little scientific evidences. Thus, in the present study, to elucidate the antitumor mechanism of SLH: in vitro and in vivo experiments were performed with hexane fraction of Solanum lyratum herba (HSLH).


METHODS
Cytotoxicity assay, 4'-6-diamidino-2-phenylindole (DAPI) staining, flow cytometric analysis for sub-G1 peaks, Western blot analysis were used with the antibodies of apoptosis related proteins in vitro. In addition, the effect of HSLH on in vivo tumor growth was evaluated in Lewis lung carcinoma (LLC) tumor model and immunohistochemistry also was performed with terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining in tumor section.


RESULTS
HSLH exhibited cytotoxicity against LLC cells most effectively among its solvent fractions. Ladder like DNA fragmentation and apoptotic features such as chromatin condensation and apoptotic bodies were observed in HSLH treated LLC cells by 4'-6-diamidino-2-phenylindole staining. HSLH also significantly increased sub-G(1) peaks, activated caspase-8, -9 and -3 proteins and cleaved poly(ADP-ribose) polymerase (PARP). Furthermore, HSLH increased the phosphorylation of extracellular signal-regulated kinase (ERK), transiently activated phospho-JNK (c-jun N-terminal kinase) and downregulated phospho-p38 MAPK. In addition, we have found for the first time HSLH treatment effectively suppressed the in vivo growth of LLC to up to approximately 30% of untreated control at 50mg/kg and significantly increased apoptotic expression in tumor section by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. Taken together, these findings strongly demonstrate that hexane fraction of Solanum lyratum herba exerts antitumor activity via caspase activation and MAPK regulation and can be effectively applied to lung cancer as a cancer chemopreventive agent.
Lee
Jin-Hyuk
JH
Cancer Preventive Material Development Research Center, College of Oriental Medicine, Kyunghee University, Seoul 130-701, Republic of Korea.
Lee
Yun-Hee
YH
Lee
Hyo-Jung
HJ
Lee
Hyo-Jeong
HJ
Lee
Eun-Ok
EO
Ahn
Kwang Seok
KS
Shim
Bum Sang
BS
Bae
Hyunsu
H
Choi
Seung-Hoon
SH
Ahn
Kyoo-Seok
KS
Baek
Nam-In
NI
Kim
Dae-Keun
DK
Kim
Sung-Hoon
SH
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D020169:Caspases / Q000378:metabolism*; D002460:Cell Line; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D020287:In Situ Nick-End Labeling; D020935:MAP Kinase Signaling System; D051379:Mice; D010936:Plant Extracts / Q000494:pharmacology*; D032322:Solanum / Q000737:chemistry*
D005456:Fluorescent Dyes; D010936:Plant Extracts; D020169:Caspases