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

本研究采用碳酸酐酶相关蛋白（carbonic anhydrase-related protein，CA-RP）表达上调和下调技术，探讨CA-RP在胃肠道肿瘤细胞增殖中的分子功能，并获得以下结果. MTT、集落形成和趋化小室分析显示，与野生型LoVo细胞相比，通过cDNA转染在培养的结肠细胞（LoVo）中异位表达CA-RP VIII增强了增殖和侵袭能力2。小干扰RNA介导的CA-RP VIII的敲低揭示了结肠癌细胞系HCT 116的细胞增殖和集落形成的显著抑制，该结肠癌细胞系显示出CA-RP VIII的高内源性表达。MTT法和趋化小室法显示，与野生型胃肠道基质细胞（GIST-T1）相比，CA-RP Xi异位表达的GIST-T1细胞增殖和侵袭能力增强4. CA异位表达 ...更多信息 与野生型PC-9细胞相比，通过cDNA转染培养的肺癌细胞（PC-9）中的-RP VIII增强了侵袭能力，这通过趋化性室测定显示5。通过使用微阵列对结肠癌细胞（LoVo）中CA-RP VIII的异位表达进行的交替表达分子的筛选分析显示了几个基因的上调和下调表达。通过实时荧光定量PCR技术再次证实了一些分子的交替表达。在所研究的基因中，层粘连蛋白β3亚基的表达被CA-RP VIII的异位表达抑制（阵列分析为0.18倍，实时PCR为0.192倍）。此外，层粘连蛋白5的其他亚基α3和γ2亚基的mRNA表达也受到抑制。据报道，层粘连蛋白5的表达降低可增强癌细胞对其他组织类型的增殖和侵袭能力。总之，研究结果表明层粘连蛋白5在CA-RP VIII介导的肿瘤发生中起作用6。利用基因芯片对GIST-T1细胞中CA-RP VIII在Xi的异位表达进行筛选分析，发现有多个基因表达上调（最高5.66倍）和下调（最低0.09倍），这些结果有力地表明CA-RP（至少VIII和Xi）具有增强肿瘤细胞增殖和侵袭能力的功能。层粘连蛋白5可能在CA-RP介导的肿瘤发生中起作用。少

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