Liu Zhujun, Qin Tingting, Yuan Xiaohan, Yang Jie, Shi Wei, Zhang Xiaoling, Jia Yanan, Liu Shaochuan, Wang Jing, Li Kai
National Clinical Research Center for Cancer, Tianjin Medical University Cancer Institute and Hospital, Tianjin, China.
Key Laboratory of Cancer Prevention and Therapy, Tianjin, China.
Front Oncol. 2022 May 18;12:875888. doi: 10.3389/fonc.2022.875888. eCollection 2022.
Bevacizumab is the representative drug in antiangiogenic therapy for lung cancer. However, it induced resistance in some neoplasm. Anlotinib, a novel multi-target tyrosine kinase inhibitor which has an inhibitory action on both angiogenesis and malignancy, is possible to reverse the resistance.
Transwell migration and invasion experiments of bevacizumab with or without anlotinib were conducted to verify the activated/inhibited ability of lung adenocarcinoma cells. We sequenced A549 cells with enhanced migration and invasion abilities after bevacizumab treatment, screened out the differentially expressed gene and further confirmed by western blot and q-PCR assays. We also investigated immunohistochemical staining of tumor tissue in mice and human lung adenocarcinoma.
Bevacizumab facilitated migration and invasion of lung adenocarcinoma cells. Differentially expressed gene RGC32 was screened out. Bevacizumab upregulated the expression of RGC32, N-cadherin, and MMP2 through ERK-MAPK and PI3K-AKT pathways. Anlotinib downregulated their expression and reversed the effect of bevacizumab on A549 cells. experiments confirmed that higher-dose bevacizumab facilitated metastasis in tumor-bearing nude mice and upregulated the expression of RGC32, N-cadherin, and MMP2, whereas anlotinib abrogated its effect. Expression of both RGC32 and N-cadherin positively correlated with lymph node metastasis and stage in lung adenocarcinoma was found. Survival analysis revealed that higher expressions of RGC32 and N-cadherin were associated with poor progression-free survival and overall survival.
Bevacizumab may promote invasion and metastasis of lung adenocarcinoma cells by upregulating RGC32 through ERK-MAPK and PI3K-AKT pathways to promote epithelial-mesenchymal transition, whereas anlotinib reverses the effect. RGC32 and N-cadherin are independent prognostic factors in lung adenocarcinoma.
贝伐单抗是肺癌抗血管生成治疗的代表性药物。然而，它会在某些肿瘤中诱导耐药。安罗替尼是一种新型多靶点酪氨酸激酶抑制剂，对血管生成和恶性肿瘤均有抑制作用，有可能逆转耐药。
进行了有或无安罗替尼的贝伐单抗的Transwell迁移和侵袭实验，以验证肺腺癌细胞的激活/抑制能力。我们对贝伐单抗治疗后迁移和侵袭能力增强的A549细胞进行测序，筛选出差异表达基因，并通过蛋白质免疫印迹和q-PCR分析进一步确认。我们还研究了小鼠肿瘤组织和人肺腺癌的免疫组化染色。
贝伐单抗促进肺腺癌细胞的迁移和侵袭。筛选出差异表达基因RGC32。贝伐单抗通过ERK-MAPK和PI3K-AKT途径上调RGC32、N-钙黏蛋白和MMP2的表达。安罗替尼下调它们的表达并逆转贝伐单抗对A549细胞的作用。实验证实，高剂量贝伐单抗促进荷瘤裸鼠的转移并上调RGC32、N-钙黏蛋白和MMP2的表达，而安罗替尼消除了其作用。发现RGC32和N-钙黏蛋白的表达均与肺腺癌的淋巴结转移和分期呈正相关。生存分析显示，RGC32和N-钙黏蛋白的高表达与无进展生存期和总生存期差有关。
贝伐单抗可能通过ERK-MAPK和PI3K-AKT途径上调RGC32以促进上皮-间质转化，从而促进肺腺癌细胞的侵袭和转移，而安罗替尼可逆转该作用。RGC32和N-钙黏蛋白是肺腺癌的独立预后因素。
