[摘 要] 目的：探讨颗粒酶M（GZMM）对肾透明细胞癌（ccRCC）细胞增殖、侵袭、迁移和血管生成的影响及相关分子机制。 方法: 采用TCGA数据库和免疫组化分析GZMM在ccRCC组织的表达及其与临床病理特征的相关性。采用CCK-8、Transwell、 划痕愈合及血管形成实验检测GZMM对ccRCC细胞增殖、侵袭、迁移和血管生成的影响，采用WB法检测GZMM对VEGF/ERK 信号通路的影响。结果: TCGA数据库和免疫组化分析表明，ccRCC组织中GZMM的表达升高（P < 0.01），且与Fuhrman分级和 淋巴结转移有关联（均P < 0.05）。GZMM高表达的患者预后不良（P < 0.05），且与FcerⅠ介导的MAPK激活有关联（P < 0.001）。 在ccRCC细胞中，干扰GZMM降低ccRCC细胞的增殖、侵袭和迁移能力，且抑制ERK信号通路（均P < 0.05）；过表达GZMM促进 ccRCC细胞的增殖、侵袭和迁移能力，且激活ERK信号通路（均P < 0.01）。在HUVEC中，分泌型GZMM促进HUVEC的增殖、迁 移、小管和血管形成的能力，且激活VEGF/ERK信号通路（均P < 0.05）。此外 ，U0126 抑制p-ERK、MMP2和MMP9的表达（均 P < 0.05），但不影响VEGFA和VEGFR2的表达。结论：ccRCC组织中GZMM呈高表达，且与其Fuhrman分级和淋巴结转移有关 联，GZMM通过激活VEGF/ERK信号通路促进ccRCC血管生成和侵袭。

[Abstract] Objective: To explore the effects of granzyme M (GZMM) on the proliferation, invasion, migration, and angiogenesis of clear cell renal cell carcinoma (ccRCC) cells and the underlying molecular mechanisms. Methods: TCGA database and immunohistochemistry were used to analyze the expression of GZMM in ccRCC tissues and its correlation with clinicopathological features. CCK-8, Transwell, wound healing, and angiogenesis assays were used to detect the effects of GZMM on the proliferation, invasion, migration, and angiogenesis of ccRCC cells. Weston blot assay was employed to detect the effects of GZMM on the VEGF/ ERK signaling pathway. Results: TCGA database and immunohistochemical analysis showed that the expression of GZMM in ccRCC tissues was elevated (P < 0.01), and was correlated with Fuhrman grading and lymph node metastasis (both P < 0.05). Patients with high expression of GZMM had a poor prognosis (P < 0.05), and it was associated with FcerⅠ-mediated MAPK activation (P < 0.001). In ccRCC cells, knockdown of GZMM decreased the proliferation, invasion and migration abilities and inhibited ERK signaling pathway (both P < 0.05) while overexpression of GZMM promoted the proliferation, invasion and migration abilities and activated ERK signaling pathway (both P < 0.01). In HUVECs, secreted GZMM promoted the proliferation, migration, tubule formation and angiogenesis abilities of HUVECs and activated VEGF/ERK signaling pathway (both P < 0.05). Furthermore, U0126 inhibited the expressions of p-ERK, MMP2, and MMP9 (all P < 0.05), but did not affect the expressions of VEGFA and VEGFR2. Conclusion: The expression of GZMM was elevated in ccRCC tissues and was correlated with its Fuhrman grading and lymph node metastasis. GZMM promoted the angiogenesis and invasion of ccRCC by activating VEGF/ERK signaling pathway.

[基金项目] 国家自然科学基金（No. 81772524）