Department of Medical Biology, Faculty of Medicine, Aydın Adnan Menderes University, Aytepe Campus, Aydin, Turkey.
Department of Pathology, Faculty of Medicine, Aydın Adnan Menderes University, Aytepe Campus, Aydin, Turkey.
Mol Biol Rep. 2024 Nov 4;51(1):1115. doi: 10.1007/s11033-024-10029-2.
Wip1, is a p53-dependent Ser/Thr phosphatase involved in the timely termination of DDR. The PPM1D gene encoding Wip1 is deregulated and thus gained an oncogene character in common human solid tumors and cell lines. This study assessed the oncogenic potential of the PPM1D gene in human non- Hodgkin's lymphomas (NHL), the most common hematological malignancy worldwide.
FFPE human lymphoid hyperplasia (LH) (n = 17) and NHL tumor lymph node samples (n = 65) and human NHL cell lines were used to assess the oncogenic potential of the PPM1D gene in the present study. Copy number gain and mRNA expression analysis of the PPM1D/Wip1 gene were assessed by qRT-PCR analysis. Mutational analysis of Exon 6 of the PPM1D gene was performed by PCR amplification and Sanger sequencing. Expressions of Wip1 and p53 proteins were assessed by immunohistochemistry and Western blot analysis.
We found that PPM1D gained gene copy number in NHL tumors by 0.7-8 times compared to the control (p < 0.01). Increased PPM1D/Wip1 gene copy number was associated with higher mRNA and protein expression in human NHL samples (p < 0.01). Overexpression of Wip1 in NHL tumors and NHL cell lines was associated with amplification level and was unaffected by p53 status. Furthermore, a heterozygous type insertion mutation was detected in exon 6 (c.1553_1554insA) of the PPM1D gene particularly in DLBCL samples. Wip1 may have oncogenic potential, perhaps playing a role in the onset and progression of human NHL. The possible significance of Wip1 overexpression to chemotherapy response in NHL remains an intriguing question that requires more exploration.
Wip1 是一种依赖于 p53 的丝氨酸/苏氨酸磷酸酶，参与 DDR 的适时终止。编码 Wip1 的 PPM1D 基因失调，因此在常见的人类实体瘤和细胞系中获得了癌基因特征。本研究评估了 PPM1D 基因在全球最常见的血液恶性肿瘤——人类非霍奇金淋巴瘤（NHL）中的致癌潜力。
本研究使用 FFPE 人类淋巴组织增生（LH）（n=17）和 NHL 肿瘤淋巴结样本（n=65）和人类 NHL 细胞系来评估 PPM1D 基因在 NHL 中的致癌潜力。通过 qRT-PCR 分析评估 PPM1D/Wip1 基因的拷贝数增益和 mRNA 表达分析。通过 PCR 扩增和 Sanger 测序对 PPM1D 基因外显子 6 的突变分析。通过免疫组织化学和 Western blot 分析评估 Wip1 和 p53 蛋白的表达。
我们发现，与对照相比，NHL 肿瘤中的 PPM1D 基因获得了 0.7-8 倍的基因拷贝数（p<0.01）。在人类 NHL 样本中，PPM1D/Wip1 基因拷贝数的增加与更高的 mRNA 和蛋白表达相关（p<0.01）。NHL 肿瘤和 NHL 细胞系中 Wip1 的过表达与扩增水平相关，与 p53 状态无关。此外，在 PPM1D 基因的外显子 6（c.1553_1554insA）中检测到杂合型插入突变，特别是在 DLBCL 样本中。Wip1 可能具有致癌潜力，可能在人类 NHL 的发病和进展中发挥作用。Wip1 过表达对 NHL 化疗反应的可能意义仍然是一个有趣的问题，需要进一步探讨。
