Department of Pediatrics, The Second Hospital of Shandong University, Jinan, Shandong 250033, P.R. China.
Department of Pediatrics, The Qilu Hospital of Shandong University, Jinan, Shandong 250012, P.R. China.
Mol Med Rep. 2018 Jan;17(1):2070-2076. doi: 10.3892/mmr.2017.8032. Epub 2017 Nov 13.
Serine-arginine protein kinase 1 (SRPK1) has been used as an important signal mediator, and is associated with cancer development. However, studies have yet to determine whether SRPK1 suppresses leukemia cell growth and induces apoptosis. Studies have also yet to reveal the underlying mechanisms. In the present study, the effects of downregulating SRPK1 gene expression on chronic myeloid leukemia cell lines (K562 cells) were investigated through RNA interference (RNAi) and the proliferation inhibition and apoptosis induction of SRPK1 in K562 cells were analyzed. K562 cells were transfected with two different concentrations of siRNA, and the transfection efficiency was detected via flow cytometry. The expression of SRPK1 was detected via reverse transcription‑quantitative polymerase chain reaction. K562 cell proliferation and apoptosis were analyzed using MTT and flow cytometry respectively. The roles of caspase‑3, poly (ADP‑ribose) polymerase (PARP), p53 and B-cell lymphoma (Bcl)‑2/Bcl‑2 associated X, apoptosis regulator (Bax) proteins in the apoptosis of human K562 cells were further examined through western blot analysis. The SRPK1 expression was lower in the K562 cells transfected with SRPK1‑siRNA compared with untransfected cells. The inhibition rate in the transfected groups was increased compared with the untransfected groups. Compared with control groups, the number of apoptotic cells in the SRPK1‑silenced groups increased. The number of early apoptotic cells also increased. The cleaved caspase‑3, cleaved PARP and p53 expression levels were significantly increased in the RNAi groups compared with control groups. Conversely, the Bcl‑2/Bax rate was significantly lower. In conclusion, the knockdown of the SRPK1 gene by RNAi inhibited the proliferation of K562 cells and induced their apoptosis. Apoptosis was induced by the activation of the PARP‑caspase3 pathway.
丝氨酸/精氨酸蛋白激酶 1（SRPK1）已被用作重要的信号介质，与癌症的发展有关。然而，研究尚未确定 SRPK1 是否抑制白血病细胞生长并诱导细胞凋亡。其潜在机制也尚未被揭示。在本研究中，通过 RNA 干扰（RNAi）下调 SRPK1 基因表达对慢性髓系白血病细胞系（K562 细胞）的影响进行了研究，并分析了 SRPK1 在 K562 细胞中的增殖抑制和凋亡诱导作用。用两种不同浓度的 siRNA 转染 K562 细胞，并通过流式细胞术检测转染效率。采用逆转录-定量聚合酶链反应检测 SRPK1 的表达。分别采用 MTT 和流式细胞术分析 K562 细胞的增殖和凋亡。通过 Western blot 分析进一步研究了 caspase-3、聚（ADP-核糖）聚合酶（PARP）、p53 和 B 细胞淋巴瘤（Bcl）-2/Bcl-2 相关 X，凋亡调节剂（Bax）蛋白在人 K562 细胞凋亡中的作用。与未转染细胞相比，转染 SRPK1-siRNA 的 K562 细胞中 SRPK1 的表达较低。与未转染组相比，转染组的抑制率增加。与对照组相比，沉默 SRPK1 组的凋亡细胞数量增加。早期凋亡细胞的数量也增加。与对照组相比，RNAi 组的 cleaved caspase-3、cleaved PARP 和 p53 表达水平显著升高，而 Bcl-2/Bax 比率显著降低。综上所述，RNAi 下调 SRPK1 基因抑制了 K562 细胞的增殖并诱导其凋亡。凋亡是通过 PARP-caspase3 途径的激活诱导的。
