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[Immuno-ricin A chain specific for gastric cancer: internalization and its significance]
1Laboratory of Gastroenterology, Xijing Hospital, Xi'an.
Biotinylated ricin A chain (RTA) was introduced to antigastric cancer monoclonal antibody MGb2 with chemical conjugating technique to form an immuno-ricin A chain conjugate (I-RTA) specific for gastric cancer. Its internalization in human gastric cancer cell line SGC-7901 was followed by double dynamic EM labelling procedure using streptavidin-gold and sheep anti-mouse IgG-gold probes. The effects of verapamil on I-RTA internalization routings were also observed. The results demonstrated that the main entry modality of I-RTA was non-coated microinvagination, followed by coated pits and interioration of microvilli. The internalized I-RTA was found to be quickly transported to multivesicular bodies and finally to lysosomes, where it was degraded, whilst in the non-membranous structures I-RTA was only occasionally encountered. Ultrastructurally, verapamil displayed its effects by making the internalized I-RTA stay longer in tubulovesicular structures and be delivered in a less amount and more slowly into lysosomes. Furthermore, I-RTA was increased in the non-membranous structures, and the in vitro killing effect was significantly enhanced. The present data revealed the basic cell biology process of I-RTA-cell interaction, which will provide not only an important clue to improving I-RTA targeting therapy efficiency, but also a new way for further screening targeting potentiators.
Biotinylated ricin A chain (RTA) was introduced to antigastric cancer monoclonal antibody MGb2 with chemical conjugating technique to form an immuno-ricin A chain conjugate (I-RTA) specific for gastric cancer. Its internalization in human gastric cancer cell line SGC-7901 was followed by double dynamic EM labelling procedure using streptavidin-gold and sheep anti-mouse IgG-gold probes. The effects of verapamil on I-RTA internalization routings were also observed. The results demonstrated that the main entry modality of I-RTA was non-coated microinvagination, followed by coated pits and interioration of microvilli. The internalized I-RTA was found to be quickly transported to multivesicular bodies and finally to lysosomes, where it was degraded, whilst in the non-membranous structures I-RTA was only occasionally encountered. Ultrastructurally, verapamil displayed its effects by making the internalized I-RTA stay longer in tubulovesicular structures and be delivered in a less amount and more slowly into lysosomes. Furthermore, I-RTA was increased in the non-membranous structures, and the in vitro killing effect was significantly enhanced. The present data revealed the basic cell biology process of I-RTA-cell interaction, which will provide not only an important clue to improving I-RTA targeting therapy efficiency, but also a new way for further screening targeting potentiators.
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