Functional analysis of carbonic anhydrase-related protein in the cell proliferation of gastrointestinal cancer

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The present study investigated molecular function of carbonic anhydrase-related protein (CA-RP) in the cell proliferation of gastrointestinal cancer by using up-and down-regulation techniques of CA-RP expression and then the following results were obtained.1. Ectopic expression of CA-RP VIII by cDNA transfection in the cultured colon cell (LoVo) enhanced proliferative and invasive abilities as compared to wild-type LoVo cells, which were shown by MTT, colony formation and chemotaxis chamber assays2..Small-interfering RNA-mediated knockdown of CA-RP VIII revealed significant inhibition in cell proliferation and colony formation of a colon cancer cell line HCT116,which showed high endogenous expression of CA-RP VIII.3. Ectopic expression of CA-RP XI by cDNA transfection in the cultured gastrointestinal stromal cell (GIST-T1) enhanced proliferative and invasive abilities as compared to wild-type GIST-T1cells, which were shown by MTT and chemotaxis chamber assays4. Ectopic expression of CA … More -RP VIII by cDNA transfection in the cultured lung cancer cell (PC-9) enhanced invasive ability as compared to wild-type PC-9 cells, which was shown by chemotaxis chamber assays5. Screening analysis for alternatively expressed molecules by ectopic expression of CA-RP VIII in colon cancer cell (LoVo) by using a microarray showed up-and down-regulated expression of several genes. Alternative expression of some molecules were reconfirmed by real-time PCR technique. Among the genes studied, expression of laminin β3 subunit was inhibited by the ectopic expression of CA-RP VIII (0.18 times by array analysis and 0.192 times by real-time PCR). Further, mRNA expression of other subunits of laminin 5,α3 and γ2 subunits, were also inhibited. Decreased expression of laminin 5 has been reported to enhance proliferative and invasive abilities of cancer cells with other tissue types. Taken together, the findings suggest that laminin 5 plays a role in the CA-RP VIII-mediated tumorinogenesis6. Screening analysis for alternatively expressed molecules by ectopic expression of CA-RP VIII in XI in GIST-T1 by using a microarray showed several genes which was up-regulated (maximum 5.66 times) and down-regulated (minimum 0.09).These findings strongly indicate that CA-RP (at least VIII and XI) function to enhance proliferative and invasive abilities of tumor cells. Laminin 5 possibly play a role in the CA-RP-mediated tumorinogeneis. Less