Faculty of Health Science, Tokoha University, 1-30, Mizuochi-cho, Aoi-ku, Shizuoka-shi, Shizuoka 420-0831, Japan.
Department of Biochemistry, The Nippon Dental University School of Life Dentistry at Tokyo, 1-9-20 Fujimi, Chiyoda-ku, Tokyo 102-8159, Japan.
Int J Mol Sci. 2018 Aug 23;19(9):2488. doi: 10.3390/ijms19092488.
Enhanced microtubule acetylation has been identified as a negative prognostic indicator in breast cancer. We reported previously that primary cultured human mammary epithelial cells manifest breast cancer-related aneuploidization via the activation of severing protein katanin-like (KL)1 when tau is deficient. To address in this current study whether microtubule hyperacetylation is involved in breast carcinogenesis through mitosis, the effects of tubacin on human mammary epithelial cells were tested using immunofluorescence techniques. Tau-knockdown cells showed enhancement of KL1-dependent events, chromosome-bridging and micronucleation in response to tubacin. These enhancements were suppressed by further expression of an acetylation-deficient tubulin mutant. Consistently, using a rat fibroblast-based microtubule sensitivity test, it was confirmed that KL1 also shows enhanced activity in response to microtubule hyperacetylation as well as katanin. It was further observed in rat fibroblasts that exogenously expressed KL1 results in more micronucleation under microtubule hyperacetylation conditions. These data suggest that microtubule acetylation upregulates KL1 and induces more aneuploidy if tau is deficient. It is thus plausible that microtubule hyperacetylation promotes tumor progression by enhancing microtubule sensitivity to KL1, thereby disrupting spindle microtubules and this process could be reversed by the microtubule-binding and microtubule protective octapeptide NAPVSIPQ (NAP) which recruits tau to the microtubules.
增强的微管乙酰化已被确定为乳腺癌的一个负预后指标。我们之前曾报道,当微管相关蛋白 tau 缺失时,原代培养的人乳腺上皮细胞通过激活丝氨酸蛋白酶 katanin-like (KL)1 表现出与乳腺癌相关的非整倍体。为了在本研究中确定微管过度乙酰化是否通过有丝分裂参与乳腺癌发生,我们使用免疫荧光技术测试了 tubacin 对人乳腺上皮细胞的影响。tau 敲低细胞表现出 KL1 依赖性事件增强,如对 tubacin 的染色体桥接和微核形成。这些增强作用被进一步表达的乙酰化缺陷型微管蛋白突变体所抑制。一致地,使用基于大鼠成纤维细胞的微管敏感性测试,证实 KL1 也表现出对微管过度乙酰化以及 katain 的增强活性。进一步在大鼠成纤维细胞中观察到,外源性表达的 KL1 在微管过度乙酰化条件下导致更多的微核形成。这些数据表明,如果 tau 缺失,微管乙酰化会上调 KL1 并诱导更多的非整倍体。因此,微管过度乙酰化通过增强微管对 KL1 的敏感性来促进肿瘤进展,从而破坏纺锤体微管,而这个过程可以通过微管结合和微管保护八肽 NAPVSIPQ (NAP) 逆转,该八肽将 tau 招募到微管上。