Diagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Meshkinfam Ave, Shiraz, Iran.
Clinical Neurology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
Biol Res. 2022 Jan 11;55(1):2. doi: 10.1186/s40659-021-00369-9.
Chinese hamster ovary cell line has been used routinely as a bioproduction factory of numerous biopharmaceuticals. So far, various engineering strategies have been recruited to improve the production efficiency of this cell line such as apoptosis engineering. Previously, it is reported that the caspase-7 deficiency in CHO cells reduces the cell proliferation rate. But the effect of this reduction on the CHO cell productivity remained unclear. Hence, in the study at hand the effect of caspase-7 deficiency was assessed on the cell growth, viability and protein expression. In addition, the enzymatic activity of caspase-3 was investigated in the absence of caspase-7.
Findings showed that in the absence of caspase-7, both cell growth and cell viability were decreased. Cell cycle analysis illustrated that the CHO knockout (CHO-KO) cells experienced a cell cycle arrest in G2/M phase. This cell cycle arrest resulted in a 1.7-fold increase in the expression of luciferase in CHO-KO cells compared to parenteral cells. Furthermore, in the apoptotic situation the enzymatic activity of caspase-3 in CHO-KO cells was approximately 3 times more than CHO-K1 cells.
These findings represented that; however, caspase-7 deficiency reduces the cell proliferation rate but the resulted cell cycle arrest leads to the enhancement of recombinant protein expression. Moreover, increasing in the caspase-3 enzymatic activity compensates the absence of caspase-7 in the caspase cascade of apoptosis.
中国仓鼠卵巢细胞系已被常规用作许多生物制药的生物生产工厂。到目前为止，已经采用了各种工程策略来提高该细胞系的生产效率，例如凋亡工程。以前有报道称，CHO 细胞中 caspase-7 的缺乏会降低细胞增殖率。但是，这种降低对 CHO 细胞生产力的影响尚不清楚。因此，在目前的研究中，评估了 caspase-7 缺乏对细胞生长、活力和蛋白表达的影响。此外，还研究了 caspase-7 缺乏时 caspase-3 的酶活性。
研究结果表明，caspase-7 缺乏时，细胞生长和细胞活力均降低。细胞周期分析表明，CHO 敲除（CHO-KO）细胞在 G2/M 期经历细胞周期停滞。与亲本细胞相比，CHO-KO 细胞中的 luciferase 表达增加了 1.7 倍。此外，在凋亡情况下，CHO-KO 细胞中的 caspase-3 酶活性比 CHO-K1 细胞高约 3 倍。
这些发现表明，caspase-7 缺乏虽然降低了细胞增殖率，但导致的细胞周期停滞导致重组蛋白表达增强。此外，caspase-3 酶活性的增加补偿了凋亡级联中 caspase-7 的缺乏。
