Purification of lectin from larvae of the fly, Musca domestica, and in vitro anti-tumor activity in MCF-7 cells
Ministry of Education, Key Laboratory of Food Nutrition and Safety, School of Food Science and Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, P. R. China
Abstract
A new lectin was purified from larvae of the fly, Musca domestica L. (Diptera: Muscidae) (MLL- 2, 38 kDa) using affinity chromatography and HPLC. Anti-tumor activity of MLL-2 was demonstrated by its inhibition of proliferation of human breast cancer (MCF-7) cells in a timeand dose-dependent manner. The results of acridine orange staining indicated that MLL-2 caused apoptosis in MCF-7 cells. DNA fragmentation in MCF-7 cells has been detected by TUNEL. Flow cytometric analysis also demonstrated that MLL-2 caused dose-dependent apoptosis of MCF-7 cells through cell arrest at G2/M phase. The MLL-2 induced a sustained increase in concentration of intracellular free calcium. Western blot revealed that MLL-2 induced apoptosis in MCF-7 cells was associated with typical apoptosis proteins in the mitochondrial pathway. In addition, the caspase-3 activity in MCF-7 cells treated with MLL-2 for 48 hours was significantly increased compared to controls (407.4 ± 3.0 vs. 1749.2 ± 6.0, P <0.01). Since MLL-2 induced apoptosis in MCF-7cells the mitochondrial pathway may be the main pathway of antitumor activity.
Keywords: Musca domestica larvae, Lectin, Apoptosis, Anti-tumor activity
Abbreviations: AO, acridine orange staining; HLF, human lung fibroblasts; MTT, growth inhibition assay; TUNEL, apoptosis assay
Correspondence:
a*caoxh@tust.edu.cn,
brubyrain121118@hotmail.com,
clumf@tust.edu.cn,
dyonghengwoxin_0824@hotmail.com,
ehelloxj2000@163.com,
fhelloxj2000@163.com,
gwangchunling@tust.edu.cn,
hzengbin@gmail.com, *Corresponding author
Associate Editor: Susan Paskewitz was editor of this paper.
Received: 26 September 2008 | Accepted: 12 January 2009 | Published: 29 September 2010
Copyright: This is an open access paper. We use the Creative Commons Attribution 3.0 license that permits unrestricted use, provided that the paper is properly attributed.
ISSN: 1536-2442 | Volume 10, Number 164
Cao X, Huo Z, Lu M, Mao D, Zhao Q, Xu C, Wang C, Zeng B. 2010. Purification of lectin from larvae of the fly, Musca domestica, and in vitro anti-tumor activity in MCF-7 cells. Journal of Insect Science 10:164, available online: insectscience.org/10.164
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