Yan Dawei, Jiang Yayi, Hu Youpeng, Hu Zhipeng, Feng Haoyue, Yue Rensong
Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, China.
Mianyang Orthopaedic Hospital, Mianyang, China.
Recent Pat Anticancer Drug Discov. 2025 Jul 3. doi: 10.2174/0115748928379439250621171215.
Pancreatic cancer (PC) is a highly aggressive malignancy with limited treatment options and poor prognosis. Dahuang Zhechong Pill, a traditional Chinese medicine, has shown promise in inhibiting inflammation. This study investigates the effects of combining Dahuang Zhechong Pill with TNS4 silencing on PC, focusing on their combined role in suppressing PC progression and exploring potential therapeutic applications. Intellectual property implications related to this combination are also explored.
Bioinformatic analysis was used to identify the key role of TNS4 in PC. CFPAC-1 PC cells were cultured and genetically modified using lentiviral transfection to stably knock down TNS4 expression. Cell proliferation was assessed using the CCK-8 assay, while cell migration and invasion capabilities were evaluated through Transwell assays. Colony formation and flow cytometry were performed to analyze clonogenic potential and cell cycle distribution, respectively. Apoptosis was assessed using tunel staining. Subcutaneous and orthotopic tumor models were established in nude mice to investigate the in vivo effects. Mice were treated with Dahuang Zhechong Pill by oral gavage. Immunohistochemistry and immunofluorescence were employed to detect the expression of key proteins involved in the NF-κB/VEGF pathway, including E-cadherin and Vimentin. ELISA was used to measure circulating IL-17 and amylase levels in mouse serum to test inflammation response.
TNS4 was upregulated in PC and positively associated with PC progression. TNS4 silencing significantly reduced CFPAC-1 cell proliferation, migration, and invasion in vitro. Flow cytometry demonstrated an increase in G0/G1 phase arrest and apoptosis in the TNS4 silencing group. Subcutaneous models showed the anti-tumor effect of TNS4 silencing. Furthermore, Dahuang Zhechong Pill treatment, when combined with TNS4 knockdown, resulted in a marked decrease in tumor size in orthotopic models. Immunohistochemical analysis revealed reduced expression of NF-κB and VEGF in tumor tissues from the combination treatment group. ELISA results indicated lower levels of serum IL-17, and amylase and higher levels of insulin in combination-treated mice.
We proposed an innovative therapeutic approach combining traditional Chinese medicine with targeted gene silencing to inhibit PC progression. By investigating the synergistic effects of TNS4 silencing and Dahuang Zhechong Pill in suppressing the NF-κB/VEGF signaling pathway, our findings highlighted a promising strategy that targets tumor proliferation and modulates the inflammatory microenvironment in PC.
Dahuang Zhechong Pill, in combination with TNS4 silencing, effectively inhibits the NF-κB/VEGF pathway and inflammation response, leading to reduced PC progression. These findings suggest a potential therapeutic approach for targeting PC through the combined use of traditional Chinese medicine and gene editing.
胰腺癌（PC）是一种侵袭性很强的恶性肿瘤，治疗选择有限且预后较差。中药大黄蛰虫丸在抑制炎症方面已显示出前景。本研究调查大黄蛰虫丸与TNS4基因沉默联合应用对胰腺癌的影响，重点关注它们在抑制胰腺癌进展中的联合作用，并探索潜在的治疗应用。还探讨了与这种联合应用相关的知识产权问题。
利用生物信息学分析确定TNS4在胰腺癌中的关键作用。培养CFPAC-1胰腺癌细胞，并使用慢病毒转染进行基因改造，以稳定敲低TNS4表达。使用CCK-8法评估细胞增殖，通过Transwell实验评估细胞迁移和侵袭能力。分别进行集落形成实验和流式细胞术分析克隆形成潜力和细胞周期分布。使用TUNEL染色评估细胞凋亡。在裸鼠中建立皮下和原位肿瘤模型以研究体内效应。通过口服灌胃给予小鼠大黄蛰虫丸。采用免疫组织化学和免疫荧光检测NF-κB/VEGF通路中关键蛋白的表达，包括E-钙黏蛋白和波形蛋白。使用ELISA检测小鼠血清中循环IL-17和淀粉酶水平以测试炎症反应。
TNS4在胰腺癌中上调，并与胰腺癌进展呈正相关。TNS4基因沉默显著降低了CFPAC-1细胞在体外的增殖、迁移和侵袭能力。流式细胞术显示TNS4基因沉默组G0/G1期阻滞增加且细胞凋亡增加。皮下模型显示了TNS4基因沉默的抗肿瘤作用。此外，大黄蛰虫丸治疗与TNS4基因敲低联合应用时，原位模型中的肿瘤大小显著减小。免疫组织化学分析显示联合治疗组肿瘤组织中NF-κB和VEGF表达降低。ELISA结果表明联合治疗小鼠血清中IL-17、淀粉酶水平较低，胰岛素水平较高。
我们提出了一种将中药与靶向基因沉默相结合以抑制胰腺癌进展的创新治疗方法。通过研究TNS4基因沉默和大黄蛰虫丸在抑制NF-κB/VEGF信号通路中的协同作用，我们的发现突出了一种有前景的策略，该策略靶向肿瘤增殖并调节胰腺癌中的炎症微环境。
大黄蛰虫丸与TNS4基因沉默联合应用可有效抑制NF-κB/VEGF通路和炎症反应，从而减少胰腺癌进展。这些发现提示了一种通过联合使用中药和基因编辑来靶向治疗胰腺癌的潜在方法。
