Anti-tumour effect of lapatinib in canine transitional cell carcinoma cell lines
Anti-tumour effect of lapatinib in canine transitional cell carcinoma cell lines
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DOI：
10.1111/vco.12434
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发表时间：
2018-12-01
影响因子：
2.1
通讯作者：
Matsuki, Naoaki
中科院分区：
文献类型：
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作者：
Sakai, Kosei;Maeda, Shingo;Matsuki, Naoaki
移行细胞癌(TCC)占发生在膀胱的犬类恶性肿瘤的90%，预后不良。我们以前的研究，使用RNA测序，表明人表皮生长因子2(HER2)是与犬TCC癌变相关的最激活的上游调控因子。本研究的目的是检测HER2酪氨酸激酶抑制剂拉帕替尼对犬TCC细胞株的体内外抗肿瘤作用。选用5种犬TCC细胞系(TCCUB、Love、SORA、LCTCC和MCTCC)。Western blotting显示HER2蛋白在所有犬TCC细胞系中均有表达。拉帕替尼以剂量依赖的方式抑制HER2的磷酸化和细胞生长。流式细胞仪细胞周期分析显示，拉帕替尼可显著增加SORA和TCCUB细胞的亚G(1)和G(0)/G(1)期细胞比例，显著降低S和G(2)/M期细胞比例。在体内实验中，将犬TCC细胞(SORA)皮下注射到裸鼠体内。接种后6天，给予拉帕替尼(100 mg/kg)或赋形剂，每日1次，连续14天。与赋形剂对照组相比，拉帕替尼组的肿瘤体积明显较小。组织学上，拉帕替尼显著增加了肿瘤组织的坏死区。这些发现表明，拉帕替尼通过抑制HER2信号转导和诱导细胞周期停滞而对犬TCC细胞产生抗肿瘤作用。
Transitional cell carcinoma (TCC) accounts for >90% of canine malignant tumours occurring in urinary bladder, and the prognosis is poor. Our previous study, using RNA sequencing, showed that human epidermal growth factor 2 (HER2) was the most activated upstream regulator related to carcinogenesis in canine TCC. The aim of this study was to examine the anti-tumour effect of lapatinib, a tyrosine kinase inhibitor of HER2, on canine TCC cell lines in vitro and in vivo. Five canine TCC cell lines (TCCUB, Love, Sora, LCTCC, and MCTCC) were used. Western blotting showed that HER2 protein expression was observed in all of the canine TCC cell lines. Lapatinib inhibited phosphorylation of HER2 and cell growth in a dose-dependent manner. Cell cycle analyses using flow cytometry showed that lapatinib significantly increased the sub-G(1) and G(0)/G(1) phase fractions and significantly decreased the S and G(2)/M phase fractions in the cell lines (Sora and TCCUB). For the in vivo experiments, the canine TCC cells (Sora) were subcutaneously injected into nude mice. Six days after inoculation, lapatinib (100 mg/kg) or vehicle was administered daily via intraperitoneal administration for 14 days. Tumour volume was significantly smaller in the lapatinib group compared with the vehicle control group. Histologically, lapatinib significantly increased necrotic areas in the tumour tissues. These findings suggest that lapatinib exerts anti-tumour effects on canine TCC cells by inhibiting HER2 signalling and inducing cell cycle arrest.