The expression level of surface marker Trop2 in gastric cancer stem-like cells
  
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DOI:10.46701/APJBG.20170217031
KeyWord:gastric cancer, CSCs, trop2, MGC803, expression
                    
AuthorInstitution
Qi Tang Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing, Jiangsu 211166, China.
Xingwang Kuai Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing, Jiangsu 211166, China.
Wei Zhao Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing, Jiangsu 211166, China.
Guipeng Ding Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing, Jiangsu 211166, China.
Zhenning Qiu Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing, Jiangsu 211166, China.
Zhuming Wang Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing, Jiangsu 211166, China.
Xiang Zou The Hong Kong Polytechnic University, Hong Kong, China.
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Abstract:
      This study aims to prepare gastric cancer stem-like cells(GCSCs) using a serum-free suspension culture, then identify it preliminarily, and observe the expression level of Trop2. Serum-free DMEM/F12 medium and low-adhesion dish were used for suspended culture of gastric cancer (GC) cell lines MGC803. The morphological characteristics of cell spheres were observed by optical microscope; the positive rates of the CD44, CD54, EpCAM and Trop2 in MGC803 and MGC803-spheres were detected by FACS; the changes of the cell cycle in the MGC803-spheres were explored by FACS and compared with that in MGC803; the mRNA level of cancer stem cells(CSCs) regulatory genes and Trop2 in MGC803-spheres and MGC803 were detected by qRT-PCR. MGC803-spheres formed after MGC803 was cultured in serum-free medium for about 14 days; the levels of CD44, CD54, EpCAM and Trop2 in MGC803-spheres group were higher than those in MGC803 group (P<0.05). The G1 phase of cell cycle in the MGC803-spheres group was obviously lower than that in MGC803 group, and the S phase of the cell cycle in MGC803-spheres group was obviously higher than that in the MGC803 group (P<0.05). The mRNA level in the CSCs regulatory genes (Oct4, Snail, Nanog) and Trop2 in MGC803-spheres group were much higher than those in MGC803 group (P<0.05). MGC803-spheres could form in serum-free and suspension culture condition. The type of cells has the characteristic of CSCs, which appears in a higher proliferation state and over expression CSCs regulating genes and Trop2.
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