Ma Zhenling, Han Xulu, Zhu Yiyang, Fu Panpan, Li Zhenzhen, Zhao Yongqin, Yang Junzheng, Liu Wei
College of Life Sciences, Henan Agricultural University, Zhengzhou, 450002, China.
Guangdong Nephrotic Drug Engineering Technology Research Center, Institute of Consun Co. for Chinese Medicine in Kidney Diseases, Guangzhou, 510530, China.
Protein J. 2025 Jun 2. doi: 10.1007/s10930-025-10273-w.
Stem cell factor (SCF) is a type of hematopoietic cytokine that functions in early hematopoietic stem cells. SCF, together with its cognate receptor c-kit, plays crucial roles in cell survival, proliferation, and differentiation. A dysregulated SCF/c-kit system is reportedly, associated strongly with various kinds of cancer in humans. However, the function and mechanism of SCF in non-small cell lung cancer (NSCLC) remains to be elucidated to date. In this context, the present study attempted to purify two different sizes of human SCF proteins and then investigate the effect of these exogenous SCFs in A549 cells. First, the homo sapiens Kit ligand (KITG) transcript variants a and b were amplified, and recombinant pET-30a vectors were constructed. Soluble His-SCF and His-SCF-2 were then expressed and purified using 0.1 mM isopropyl-β-D-1-tiogalactopiranoside (IPTG) at 25 °C and 0.3 mM IPTG at 25 °C, respectively. MTT assay was conducted next, which revealed that the purified proteins significantly promoted A549 cell activity and increased the phosphorylation of P65. Further, NF-κB pathway blockade was achieved through treatment with BAY11-7082, and it was observed to decrease the A549 cell activity promoted by the purified proteins. These findings provided insights into the biological role of SCF in the development of NSCLC.
干细胞因子（SCF）是一种在早期造血干细胞中起作用的造血细胞因子。SCF与其同源受体c-kit一起，在细胞存活、增殖和分化中发挥关键作用。据报道，失调的SCF/c-kit系统与人类的各种癌症密切相关。然而，迄今为止，SCF在非小细胞肺癌（NSCLC）中的功能和机制仍有待阐明。在此背景下，本研究试图纯化两种不同大小的人SCF蛋白，然后研究这些外源性SCF对A549细胞的影响。首先，扩增智人Kit配体（KITG）转录变体a和b，并构建重组pET-30a载体。然后分别在25℃使用0.1 mM异丙基-β-D-1-硫代半乳糖苷（IPTG）和在25℃使用0.3 mM IPTG表达并纯化可溶性His-SCF和His-SCF-2。接下来进行MTT试验，结果显示纯化的蛋白显著促进A549细胞活性并增加P65的磷酸化。此外，通过用BAY11-7082处理实现了NF-κB途径的阻断，并且观察到其降低了纯化蛋白促进的A549细胞活性。这些发现为SCF在NSCLC发生发展中的生物学作用提供了见解。
