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目的 探究硼替佐米在抑制胰腺癌细胞恶性生物学行为和肝转移方面的潜能。方法 以KPC和Panc02细胞为研究对象，CCK-8实验检测硼替佐米对胰腺癌细胞活力的影响，并计算半抑制浓度(IC50)值；通过EdU染色检测硼替佐米对胰腺癌细胞活力的影响；凋亡实验用于检测硼替佐米对胰腺癌细胞的促凋亡作用；集落形成、细胞划痕和Transwell实验检测硼替佐米对胰腺癌细胞集落形成、迁移和侵袭能力的影响；qRT-PCR和Western blot用于检测硼替佐米对细胞上皮间质转化(EMT)相关标志物分子，包括Cdh1、Cdh2、Vim和Snail基因，E-cadherin蛋白(上皮钙黏蛋白)、神经钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)和Snail蛋白表达的影响；动物脾转肝模型用于评估硼替佐米对于抑制胰腺癌肝转移的治疗价值。结果 CCK-8实验显示硼替佐米能降低胰腺癌KPC和Panc02细胞的活力(P<0.000 1),IC50值分别约为118.70 nmol/L和34.16 nmol/L;EdU实验显示硼替佐米能抑制胰腺癌细胞的增殖能力(P<0.01);凋亡实验显示硼替佐米能促进胰腺癌细胞发生早期凋亡和晚期凋亡(P<0.05);集落形成、细胞划痕和Transwell实验显示硼替佐米能抑制胰腺癌细胞的集落形成、迁移和侵袭能力(P<0.01);qRT-PCR和Western blot实验显示硼替佐米能够改变胰腺癌细胞EMT相关标志物分子的mRNA(P<0.000 1)(Cdh1基因表达水平提高，Cdh2、Vim和Snail基因表达水平降低)和蛋白(E-cadherin蛋白水平提高，N-cadherin、Vimentin和Snail蛋白水平降低)表达；动物脾转肝模型表明硼替佐米能够有效限制胰腺癌肝转移能力(P<0.01)。结论 硼替佐米能够抑制胰腺癌细胞多种恶性生物学行为和肝转移，可能是胰腺癌治疗的潜在药物。
Abstract：Objective To investigate the potential of bortezomib in inhibiting the malignant biological behaviors and liver metastasis of pancreatic cancer cells. Methods KPC and Panc02 cell lines were used as a couple of working cells in this study. The CCK-8 assay was used to assess the effect of bortezomib on cell viability in murine pancreatic cancer cells, and the half inhibitory concentration(IC50) values were calculated. The EdU assay was performed to evaluate the impact of bortezomib on the proliferation of these two cell lines. Apoptosis assays were conducted to investigate the pro-apoptotic effects of bortezomib on pancreatic cancer cells. Colony formation, scratch wound healing, and Transwell assays were used to examine the effects of bortezomib on the proliferation, migration, and invasion of pancreatic cancer cells. QRT-PCR and Western blot were employed to assess the impact of bortezomib on the expression of epithelial-mesenchymal transition(EMT) related markers, including Cdh1, Cdh2, Vim and Snail gene and E-cadherin protein, N-cadherin protein, Vimentin protein and Snail protein. An in vivo spleen-to-liver metastasis model was established to evaluate the therapeutic value of bortezomib in inhibiting pancreatic cancer liver metastasis. Results The CCK-8 assay showed that bortezomib significantly reduced the viability of KPC and Panc02 cells(P<0.000 1), with IC50 values of approximately 118.70 nmol/L and 34.16 nmol/L, respectively. The EdU assay showed that bortezomib markedly inhibited the proliferative capacity of pancreatic cancer cells(P<0.01). Apoptosis assays showed that bortezomib promoted both early and late apoptosis in pancreatic cancer cells(P<0.05). Colony formation, scratch wound healing, and Transwell assays showed that bortezomib effectively suppressed colony formation, migration, and invasion(P<0.01) of pancreatic cancer cells. qRT-PCR and Western blot analyses showed that bortezomib altered the expression of EMT-related markers at both the mRNA(the expression level of Cdh1 gene increased, while the expression levels of Cdh2, Vim and Snail genes decreased)(P<0.000 1) and protein(the expression level of E-cadherin protein increased, while the expression levels of N-cadherin protein, Vimentin protein and Snail protein decreased) levels in pancreatic cancer cells. The in vivo spleen-to-liver metastasis model demonstrated that bortezomib significantly inhibited pancreatic cancer liver metastasis(P<0.01). Conclusion Bortezomib can inhibit the malignant biological behaviors of pancreatic cancer cells, suggesting it might be a potential anti-cancer drug in pancreatic cancer.
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基本信息:
DOI：10.19405/j.cnki.issn1000-1492.2025.05.008
中图分类号:R735.9
引用信息:
[1]陈崟峰,黄星.硼替佐米抑制胰腺癌细胞恶性生物学行为和肝转移[J].安徽医科大学学报,2025,60(05):825-833.DOI:10.19405/j.cnki.issn1000-1492.2025.05.008.
基金信息:
浙江省自然科学基金杰出青年基金项目(编号：LR22H160010)~~