SUZ12 inhibition attenuates cell proliferation of glioblastoma via post-translational regulation of CDKN1B,Genes & Genomics

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SUZ12 inhibition attenuates cell proliferation of glioblastoma via post-translational regulation of CDKN1B
Genes & Genomics ( IF 1.6 ) Pub Date : 2023-10-19 , DOI: 10.1007/s13258-023-01468-5
Sojin Kim ₁ , Sungsin Jo ₂ , Sun Ha Paek ₃ , Sang Soo Kang ₄ , Heekyoung Chung _{5,

6}

Affiliation

Background

Human gliomas are aggressive brain tumors characterized by uncontrolled cell proliferation. Differential expression of Polycomb repressive complex 2 (PRC2) has been reported in various subtypes of glioma. However, the role of PRC2 in uncontrolled growth in glioma and its underlying molecular mechanisms remain to be elucidated.

Objective

We aimed to investigate the functional role of PRC2 in human glioblastoma cell growth by silencing SUZ12, the non-catalytic core component of PRC2.

Methods

Knockdown of SUZ12 was achieved by infecting T98G cells with lentivirus carrying sequences specifically targeting SUZ12 (shSUZ12). Gene expression was examined by quantitative PCR and western analysis. The impact of shSUZ12 on cell growth was assessed using a cell proliferation assay. Cell cycle distribution was analyzed by flow cytometry, and protein stability was evaluated in cycloheximide-treated cells. Subcellular localization was examined through immunofluorescence staining and biochemical cytoplasmic-nuclear fractionation. Gene expression analysis was also performed on human specimens from normal brain and glioblastoma patients.

Results

SUZ12 knockdown (SUZ12 KD) led to widespread decrease in the PRC2-specific histone mark, accompanied by a slowdown of cell proliferation through G1 arrest. In SUZ12 KD cells, the degradation of CDKN1B protein was reduced, resulting from alterations in the MYC-SKP2-CDKN1B axis. Furthermore, nuclear localization of CDKN1B was enhanced in SUZ12 KD cells. Analysis of human glioblastoma samples yielded increased expression of EZH2 and MYC along with reduced CDKN1B compared to normal human brain tissue.

Conclusion

Our findings suggest a novel role for SUZ12 in cell proliferation through post-translational regulation of CDKN1B in glioblastoma.

中文翻译：

SUZ12 抑制通过 CDKN1B 的翻译后调节减弱胶质母细胞瘤的细胞增殖

背景

人类神经胶质瘤是一种侵袭性脑肿瘤，其特征是细胞增殖不受控制。据报道，Polycomb 抑制复合物 2 (PRC2) 在各种神经胶质瘤亚型中存在差异表达。然而，PRC2 在神经胶质瘤失控生长中的作用及其潜在的分子机制仍有待阐明。

客观的

我们旨在通过沉默 PRC2 的非催化核心成分 SUZ12 来研究 PRC2 在人胶质母细胞瘤细胞生长中的功能作用。

方法

SUZ12的敲低是通过用携带特异性靶向SUZ12 (shSUZ12) 的序列的慢病毒感染 T98G 细胞来实现的。通过定量PCR和蛋白质印迹分析检查基因表达。使用细胞增殖测定评估 shSUZ12 对细胞生长的影响。通过流式细胞术分析细胞周期分布，并评估放线菌酮处理的细胞中的蛋白质稳定性。通过免疫荧光染色和生化细胞质-核分级分离检查亚细胞定位。还对正常大脑和胶质母细胞瘤患者的人类样本进行了基因表达分析。

结果

SUZ12 敲低 (SUZ12 KD) 导致 PRC2 特异性组蛋白标记广泛减少，并伴随着 G1 期停滞导致的细胞增殖减慢。在 SUZ12 KD 细胞中，由于 MYC-SKP2-CDKN1B 轴的改变，CDKN1B 蛋白的降解减少。此外，SUZ12 KD 细胞中 CDKN1B 的核定位得到增强。对人胶质母细胞瘤样本的分析发现，与正常人脑组织相比，EZH2 和 MYC 的表达增加，而 CDKN1B 的表达减少。