Identification of proteins of human colorectal carcinoma cell line SW480 by two-dimensional electrophoresis and MALDI-TOF mass spectrometry

AIM： To conduct bhe proteomic analysis of human colorectal carcinoma cell line, SW480 by using two-dimensional electrophoresis （2-DE） and matrix-assisted laser desorption /ionization-time of flight mass spectrometry （MALDITOFMS）. METHODS： The total proteins of human colorectal carcinoma cell line, SW480 were separated with 2-DE by using immobilized pH gradient strips and visualized by staining with silver nitrate. The gel images were acquired by scanner and 2-DE analysis software, Image Master 2D Elite. Nineteen distinct protein spots were excised from gel randomly and digested in gel by TPCK-trypsin. Mass analysis of the byptic digest peptides mixture was performed by using MALDI-TOF MS. Peptide mass fingerprints （PMFs） obtained by the MALDI-TOF analysis were used to search NCBI, SWISS-PROT and MSDB databases by using Mascot software. RESULTS： PMF maps of all spots were obtained by MALDI-TOF MS and thirteen proteins were preliminarily identified. CONCLUSION： The methods of analysis and identification of protein spots of tumor cells in 2-DE gel with silver staining by MALDI-TOF MS derived PMF have been established. Protein expression profile of SW480 has been obtained. It is demonstrated that a combination of proteomics and cell culture is a useful approach to comprehend the process of colon carcinogenesis.

PRLs Promote Spreading, Adhesion, and Proliferation of Human SW480 and SW620 Cells

Recent studies suggest that PRL-3 is involved in the metastasis of colorectal cancer, but the mechanism concerning that has not been well defined. This article expresses PRL-1, PRL-2, and PRL-3 and the catalytically inactive mutant forms of those enzymes in SW620 and SW480 cells, two human cell lines derived from non-metastatic cancer and metastatic colorectal cancer, respectively. While the expression of the native forms of PRLs promotes the spreading, adhesion, and proliferation of these cells, the expression of their mutant forms inhibits the earlier-mentioned processes. These data thus provide a cellular mechanism for the role of PRL-3 in tumor metastasis and suggest that all the three PRLs have similar functions.

Construction of Recombinant Adenovirus Carrying GRIM19 and Its Effect on SW480 Cells

In order to examine the effect of GRIM19 on colon cancer cell SW480, the recombinant adenovirus carrying GRIM19 gene was constructed and transfected into SW480 cells. GRIM19 cDNA was amplified by PCR with the template pcxn2-GRIM19 and cloned into the shuttle plasmid pAdTrack-CMV. The plasmid pAdTrack-CMV-GRIM19 was linearized by PmeⅠ and homologously recombined with bone plasmid pAdEasy-1 in BJ5183, followed by identification by enzyme digestion. After transfection of linearized pAd-GRIM19 with PacⅠ into HEK293 cells, Ad-GRIM19 was obtained and amplified by 3 circles. SW480 cells were infected with Ad-GRIM19. The apoptosis rate was detected by flow cytometry. Agarose electrophoresis revealed the bands of recombinant plasmids identified by enzyme digestion were in the right range corresponding with expectation. Under the fluorescent microscopy, the package of Ad-GRIM19 in HEK293 cells and the expression of Ad-GRIM19 in SW480 cells were observed. The transfection of Ad-GRIM19 into SW480 cells increased the apoptosis rate of SW480 cells as compared with controls, It was concluded that Ad-GRIM19 was successfully constructed and the overexpression of GRIM19 in colon cancer cell lines could promote the apoptotic cell death.

Growth Inhibition Effect of DL-Lysine Acetylalicylate on sw480 Colon Carcinoma Cells

Objective： To investigate the effect of DL-lysine acetylsalicylate on proliferation of colon carcinoma cells line sw480. Methods： After treatment of DL-lysine acetylsalicylate, the study was performed by observing sw480 colorectal cancer cells with phase contrast microscope, making growth curve, and examining the inhibition rate of sw480 cells with MTT assay. Results： The morphology of sw480 cells showed characteristics of apoptosis, the cell growth curve showed inhibited proliferation of sw480 cells when treated with DL-lysine acetylsalicylate （P〈0.05）. The rate of inhibition was upward when the drug concentration increased. Conclusion： DL-lysine acetylsalicylate for injection can inhibit the growth of sw480 colorectal cancer cells obviously in a dose dependent manner.

Chinese Herb Formulae Inhibit the Proliferation of Human Colon Cancer SW480 Cells by Inducing Cell Apoptosis

Objective:This study aimed to reveal the antitumor effects of Chinese herbal formulae and the underlying mechanisms in treating colorectal cancer,with a focus on developing traditional Chinese medicine(TCM)as a supplement and alternative therapeutic method for cancers.Materials and Methods:Human colon cancer SW480 cells were treated with three Chinese herbal formulae,Bu Zhong Yi Qi Decoction,Fuzi Lizhong Decoction,and Pulsatilla Decoction at different concentrations(50–600μg/mL)for 24,36,and 48 h,respectively.Cell viability was determined using the resazurin reduction assay,and cell survival rate was evaluated using a colony formation assay.After treatment with different concentrations(50–600μg/mL)of these three formulae for 48 h,the effects of the Chinese herbal formulae on cell apoptosis were investigated using Hoechst/propidium iodide(PI)staining.The positive PI-stained cells were investigated using an EnSpire multilabel plate reader and the positive Hoechst-stained cells were observed under a fluorescence microscope for morphological changes.Results:Bu Zhong Yi Qi Decoction,Fuzi Lizhong Decoction,and Pulsatilla Decoction inhibited SW480 cell proliferation in a dose-and time-dependent manner and induced cell apoptosis.Conclusion:Chinese herbal formulae with a special prescription form of TCM with antitumor effects bring a new perspective in line with the principles of TCM in cancer treatment.