Biomedical Research Center, SAS, Dubravska cesta 9, 845 05 Bratislava, Slovakia.
Centre of Experimental Medicine, SAS, Dubravska cesta 9, 841 04 Bratislava, Slovakia.
Int J Mol Sci. 2020 Jan 18;21(2):639. doi: 10.3390/ijms21020639.
In our study, we performed retroviral transduction to overexpress codon-optimized variant of gene encoding human carbonic anhydrase I (optiCA1) in two tumor cell lines PC3 and MDA-MB-231, derived from human prostatic and breast carcinoma respectively. We achieved significantly enhanced and stable overexpression of exogenous optiCA1 gene. The expression of endogenous, wild CA1 gene was found to be normally low (C 28.6 for PC3 cells) or below to the detection limit (C 35.5 for MDA-MB-231 cells). No morphological changes and no decreasing viability of tumor cells were observed upon stable overexpression of the optiCA1 gene. In our study we have shown that the overexpression of the optimized human CA1 in engineered PC3 and MDA-MB-231 cells did not induce similar changes as we observed in tumor cells cultivated in the presence of human sera containing extensively high titers of anti-CA I autoantibodies from patients with complete remission of malignant disease. In both optiCA1transduced cell lines, the expression of selected genes responsible for basal lamina assembly, cytoskeleton, extracellular matrix proteins and proto-oncogenes (COL1A1, COL4A4, LAMC2, CTHRC1, and WNT7B) was not changed.
在我们的研究中，我们通过逆转录病毒转导过表达了经密码子优化的编码人碳酸酐酶 I（optiCA1）的基因变体，该基因在源自人前列腺癌和乳腺癌的两种肿瘤细胞系 PC3 和 MDA-MB-231 中表达。我们实现了外源性 optiCA1 基因的显著增强和稳定过表达。内源性野生 CA1 基因的表达被发现正常较低（PC3 细胞为 C 28.6）或低于检测限（MDA-MB-231 细胞为 C 35.5）。在稳定过表达 optiCA1 基因时，肿瘤细胞没有观察到形态变化和活力降低。在我们的研究中，我们表明在工程化的 PC3 和 MDA-MB-231 细胞中过表达优化的人 CA1 不会诱导与我们在含有恶性疾病完全缓解患者的高滴度抗 CA1 自身抗体的人血清中培养的肿瘤细胞中观察到的相似变化。在两种 optiCA1 转导的细胞系中，负责基底膜组装、细胞骨架、细胞外基质蛋白和原癌基因（COL1A1、COL4A4、LAMC2、CTHRC1 和 WNT7B）的选定基因的表达没有改变。
