Institute for Regenerative Medicine, Shanghai East Hospital, Tongji University, Shanghai, 200123, China.
Department of Hepatobiliary Surgery, Xijing Hospital, Air Force Medical University, Xi'an, Shaanxi Province, 710032, China.
Cell Death Dis. 2020 Nov 12;11(11):974. doi: 10.1038/s41419-020-03149-7.
Translationally controlled tumor protein (TCTP) is a highly conserved protein that accumulated in the tumorigenesis of various malignancies. Despite the important role of TCTP protein in tumor progression, the precise function and underlying mechanistic regulation of TCTP mRNA in hepatocellular carcinoma (HCC) remain unclear. In this study, we found that TCTP protein was overexpressed in HCC patients but TCTP mRNA expression levels were reversed. TCTP knockout HCC cells exhibited attenuated abilities of proliferation, migration, and invasion. The knockdown of TCTP by siRNA effectively reduced TCTP mRNA levels but not protein levels in HCC cells. Moreover, although the constitutive knockdown of TCTP inhibited almost 80% of TCTP protein expression levels in tumors of wildtype transgenic mice (TCTP KD/WT), partial restoration of TCTP protein expression was observed in the tumors of heterozygous TCTP mice (TCTP KD/TCTP±). The blockage of mRNA synthesis with ActD stimulated TCTP protein expression in HCC cells. In contrast, combined treatment with ActD and CHX or MG132 treatment alone did not lead to the TCTP protein accumulation in cells. Furthermore, following the introduction of exogenous TCTP in cells and orthotopic HCC tumor models, the endogenous TCTP protein did not change with the recombinational TCTP expression and kept a rather stable level. Dual-luciferase assays revealed that the coding sequence of TCTP mRNA functions as a sponge to regulate the TCTP protein expression. Collectively, our results indicated that the TCTP mRNA and protein formed a closed regulatory circuit and works as a buffering system to keep the homeostasis of TCTP protein levels in HCC.
翻译控制肿瘤蛋白(TCTP)是一种高度保守的蛋白质,在各种恶性肿瘤的发生中积累。尽管 TCTP 蛋白在肿瘤进展中具有重要作用,但 TCTP mRNA 在肝细胞癌(HCC)中的精确功能和潜在的机制调节仍不清楚。在这项研究中,我们发现 TCTP 蛋白在 HCC 患者中过度表达,但 TCTP mRNA 表达水平相反。TCTP 敲除 HCC 细胞表现出增殖、迁移和侵袭能力减弱。siRNA 下调 TCTP 有效降低了 HCC 细胞中 TCTP mRNA 的水平,但不降低 TCTP 蛋白的水平。此外,尽管 TCTP 的组成性敲低在野生型转基因小鼠(TCTP KD/WT)的肿瘤中几乎抑制了 80%的 TCTP 蛋白表达水平,但在杂合 TCTP 小鼠(TCTP KD/TCTP±)的肿瘤中观察到 TCTP 蛋白表达的部分恢复。用 ActD 阻断 mRNA 合成刺激 HCC 细胞中的 TCTP 蛋白表达。相比之下,用 ActD 和 CHX 联合处理或单独用 MG132 处理不会导致细胞中 TCTP 蛋白的积累。此外,在细胞和原位 HCC 肿瘤模型中引入外源性 TCTP 后,内源性 TCTP 蛋白不会随重组 TCTP 表达而改变,保持相当稳定的水平。双荧光素酶报告基因实验表明,TCTP mRNA 的编码序列作为一种海绵,调节 TCTP 蛋白的表达。总之,我们的结果表明,TCTP mRNA 和蛋白形成了一个封闭的调节回路,作为一个缓冲系统,维持 HCC 中 TCTP 蛋白水平的内稳态。