Calvatia lilacina protein-extract induces apoptosis through glutathione depletion in human colorectal carcinoma cells.
This paper reports that a novel protein extract isolated from Calvatia lilacina (CL) can induce cell death against four types of human colorectal cancer cells. Importantly, CL was shown to be free of apoptotic effects against normal rat liver cells. We have also identified that CL-induced glutathione (GSH) depletion is the major contributor responsible for the apoptotic cell death induction of SW 480 cells, as evidenced by the observation that exogenously added N-acetylcysteine (NAC), or GSH, but not vitamin C, could offer a near complete protection of CL-treated cells against apoptotic cell death. Furthermore, the participation of reactive oxygen species (ROS) evoked a drop in the transmembrane potential (Delta Psi(m)) in the CL-induced apoptotic cell death. This observation can only be deemed as a minor pathway due to the fact that cyclosporine A (CyA) could only partially rescue the CL-treated cells from apoptotic cell death. Likewise, despite the fact that CL could induce the upregulation of Bax, its knockdown via siRNA (48 h) failed to completely mitigate apoptotic cell death, indicating that its role in this apoptotic process was insignificant. To further explore the possible underlying mechanism associated with CL-induced GSH depletion, we proceeded to determine the effect of CL on the cellular gamma-glutamylcysteine synthetase (gamma-GCS), a rate-limiting enzyme responsible for GSH biosynthesis, and demonstrated that indeed gamma-GCS could be repressed by CL. Taken together, we report here for the first time that the anticancer effect of CL on human colorectal cancer cells is mediated through GSH depletion mechanism rather than a ROS-mediated killing process. This functional attribute of CL can thus provide the basis for the strategic design of a treatment of colorectal cancer.
Tsay
Jwu-Guh
JG
Department of Nutrition and Health Science, Toko University, Puzih City, Chiayi County 61363, Taiwan.
Chung
King-Thom
KT
Yeh
Chung-Hung
CH
Chen
Wan-Ling
WL
Chen
Chi-Hung
CH
Lin
Martin Hsiu-Chu
MH
Lu
Fung-Jou
FJ
Chiou
Jeng-Fong
JF
Chen
Ching-Hsein
CH
D000363:Agaricales / Q000737:chemistry*; D000970:Antineoplastic Agents; D017209:Apoptosis / Q000187:drug effects*; D045744:Cell Line, Tumor; D015179:Colorectal Neoplasms; D004249:DNA Damage / Q000187:drug effects; D005978:Glutathione / Q000032:analysis*; D006801:Humans; D020287:In Situ Nick-End Labeling; D008928:Mitochondria / Q000187:drug effects; D011506:Proteins / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000032:analysis
D000970:Antineoplastic Agents; D011506:Proteins; D017382:Reactive Oxygen Species; D051028:bcl-2-Associated X Protein; D005978:Glutathione