Characterization of the loss of SUMO pathway function on cancer cells and tumor proliferation.
Characterization of the loss of SUMO pathway function on cancer cells and tumor proliferation.
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DOI：
10.1371/journal.pone.0123882
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Bence N
中科院分区：
文献类型：
--
作者：
He X;Riceberg J;Pulukuri SM;Grossman S;Shinde V;Shah P;Brownell JE;Dick L;Newcomb J;Bence N
SUMO化是一种翻译后泛素样蛋白修饰途径，调节重要的细胞过程，包括染色体结构、动粒功能、染色体分离、核和亚核组织、转录和DNA损伤修复。越来越多的证据表明，SUMO通路在癌症中失调，提高了调节该通路可能具有治疗潜力的可能性。为了研究SUMO通路在癌细胞增殖和肿瘤生长中的重要性，我们应用基于慢病毒的短发夹RNA（shRNA）来敲低人类癌细胞中的SUMO通路基因。SAE 2和UBC 9的shRNA降低了SUMO缀合活性并抑制了人癌细胞的增殖。为了扩展这些观察结果，我们产生了多西环素诱导的条件性SAE 2 shRNA细胞系，以实现急性和可逆的SAE 2敲低。在U2 OS和HCT 116细胞中，条件性SAE 2敲低减缓了体外细胞生长，并且SAE 2敲低诱导了多种终末结果，包括凋亡、核内复制和衰老。多核细胞衰老，衰老标记物SA-β Gal染色阳性，p53和p21水平升高。为了解释这些表型，我们证实SUMO途径活性的丧失导致SUMO化拓扑异构酶IIα的丧失和染色质桥的出现，这可能损害正常的胞质分裂并导致多核化。此外，SAE 2的敲低诱导PML核体的破坏，这可能进一步促进细胞凋亡或衰老。在体内HCT 116异种移植肿瘤模型中，条件性SAE 2敲低强烈损害肿瘤生长。这些数据表明，SUMO通路是体外癌细胞增殖和体内肿瘤生长所必需的，暗示SUMO通路是潜在的癌症治疗靶点。
SUMOylation is a post-translational ubiquitin-like protein modification pathway that regulates important cellular processes including chromosome structure, kinetochore function, chromosome segregation, nuclear and sub-nuclear organization, transcription and DNA damage repair. There is increasing evidence that the SUMO pathway is dysregulated in cancer, raising the possibility that modulation of this pathway may have therapeutic potential. To investigate the importance of the SUMO pathway in the context of cancer cell proliferation and tumor growth, we applied lentivirus-based short hairpin RNAs (shRNA) to knockdown SUMO pathway genes in human cancer cells. shRNAs for SAE2 and UBC9 reduced SUMO conjugation activity and inhibited proliferation of human cancer cells. To expand upon these observations, we generated doxycycline inducible conditional shRNA cell lines for SAE2 to achieve acute and reversible SAE2 knockdown. Conditional SAE2 knockdown in U2OS and HCT116 cells slowed cell growth in vitro, and SAE2 knockdown induced multiple terminal outcomes including apoptosis, endoreduplication and senescence. Multinucleated cells became senescent and stained positive for the senescence marker, SA-β Gal, and displayed elevated levels of p53 and p21. In an attempt to explain these phenotypes, we confirmed that loss of SUMO pathway activity leads to a loss of SUMOylated Topoisomerase IIα and the appearance of chromatin bridges which can impair proper cytokinesis and lead to multinucleation. Furthermore, knockdown of SAE2 induces disruption of PML nuclear bodies which may further promote apoptosis or senescence. In an in vivo HCT116 xenograft tumor model, conditional SAE2 knockdown strongly impaired tumor growth. These data demonstrate that the SUMO pathway is required for cancer cell proliferation in vitro and tumor growth in vivo, implicating the SUMO pathway as a potential cancer therapeutic target.
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影响因子：
8
作者：
Bower, J. J.;Karaca, G. F.;Zhou, Y.;Simpson, D. A.;Cordeiro-Stone, M.;Kaufmann, W. K.
通讯作者：
Kaufmann, W. K.
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