4beta-Hydroxywithanolide E from Physalis peruviana (golden berry) inhibits growth of human lung cancer cells through DNA damage, apoptosis and G2/M arrest.
BACKGROUND
The crude extract of the fruit bearing plant, Physalis peruviana (golden berry), demonstrated anti-hepatoma and anti-inflammatory activities. However, the cellular mechanism involved in this process is still unknown.


METHODS
Herein, we isolated the main pure compound, 4beta-Hydroxywithanolide (4betaHWE) derived from golden berries, and investigated its antiproliferative effect on a human lung cancer cell line (H1299) using survival, cell cycle, and apoptosis analyses. An alkaline comet-nuclear extract (NE) assay was used to evaluate the DNA damage due to the drug.


RESULTS
It was shown that DNA damage was significantly induced by 1, 5, and 10 microg/mL 4betaHWE for 2 h in a dose-dependent manner (p < 0.005). A trypan blue exclusion assay showed that the proliferation of cells was inhibited by 4betaHWE in both dose- and time-dependent manners (p < 0.05 and 0.001 for 24 and 48 h, respectively). The half maximal inhibitory concentrations (IC50) of 4betaHWE in H1299 cells for 24 and 48 h were 0.6 and 0.71 microg/mL, respectively, suggesting it could be a potential therapeutic agent against lung cancer. In a flow cytometric analysis, 4betaHWE produced cell cycle perturbation in the form of sub-G1 accumulation and slight arrest at the G2/M phase with 1 microg/mL for 12 and 24 h, respectively. Using flow cytometric and annexin V/propidium iodide immunofluorescence double-staining techniques, these phenomena were proven to be apoptosis and complete G2/M arrest for H1299 cells treated with 5 microg/mL for 24 h.


CONCLUSIONS
In this study, we demonstrated that golden berry-derived 4betaHWE is a potential DNA-damaging and chemotherapeutic agent against lung cancer.
Yen
Ching-Yu
CY
Department of Oral and Maxillofacial Surgery, Chi-Mei Medical Center, Tainan, Taiwan.
Chiu
Chien-Chih
CC
Chang
Fang-Rong
FR
Chen
Jeff Yi-Fu
JY
Hwang
Chi-Ching
CC
Hseu
You-Cheng
YC
Yang
Hsin-Ling
HL
Lee
Alan Yueh-Luen
AY
Tsai
Ming-Tz
MT
Guo
Zong-Lun
ZL
Cheng
Yu-Shan
YS
Liu
Yin-Chang
YC
Lan
Yu-Hsuan
YH
Chang
Yu-Ching
YC
Ko
Ying-Chin
YC
Chang
Hsueh-Wei
HW
Wu
Yang-Chang
YC
D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis; D002455:Cell Division; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D020552:Comet Assay / Q000379:methods; D004249:DNA Damage; D004305:Dose-Response Relationship, Drug; D004354:Drug Screening Assays, Antitumor; D016195:G2 Phase; D006801:Humans; D020128:Inhibitory Concentration 50; D008175:Lung Neoplasms / Q000188:drug therapy*; D032321:Physalis / Q000378:metabolism*; D010936:Plant Extracts / Q000494:pharmacology*; D054358:Withanolides / Q000494:pharmacology*
C554450:4-hydroxywithanolide E; D000970:Antineoplastic Agents; D010936:Plant Extracts; D054358:Withanolides