Background

Lung cancer is one of the most common malignancies worldwide. To treat lung cancer, various anticancer drugs were developed and tested, but they failed because of drug resistance. In the present study, we tested herbal medicines, such as TK and CuD, as anticancer drugs to decrease side effects and resistance.

Methods

Cell viability was measured by an MTT assay. Analysis of cell cycle arrest was performed by flow cytometry. Induction of apoptosis by cucurbitacin D was measured by an annexin V-FITC/PI assay. We performed RTK kit analysis. Levels of p-ErbB3, p-STAT3, p-NF-κB, and caspases were measured by western blot analysis. Nuclear staining of ErbB3 was measured by immunocytochemistry. Transcriptional activity of STAT3 and NF-κB was detected by STAT3 and NF-κB luciferase reporter gene assays.

Results

We found a synergistic effect of TK with CDDP and PXD in primary culture of human NSCLC tumor cells. The combination of CDDP/PXD and TK or CuD inhibited the proliferation of H1299 cells. The combination of CDDP/PXD and TK or CuD induced sub-G1 and G2/M cell cycle arrest in H1299 cells. The combination of CDDP/PXD and TK or CuD induced apoptosis, regulated apoptotic molecules, caused morphological changes and inhibited colony formation in H1299 cells. We found that TK suppresses p-ErbB3 expression and signaling. The combination of CDDP/PXD and TK or CuD inhibited p-AKT, p-Erk, and p-JNK signaling and suppressed Stat3 and NF-κB transcriptional activity in H1299 cells. More importantly, the combination of CDDP/PXD and TK or CuD inhibited p-ErbB3 and downstream molecules in H1299 cells. The combination of CDDP/PXD and TK or CuD inhibited ErbB2/ErbB3 dimerization. Our results clearly demonstrate that the synergistic effect of CDDP/PXD and TK or CuD inhibits cell growth and induces apoptosis by inhibiting ErbB3 signaling.

Conclusion

The combination of CDDP/PXD and TK or CuD decreases cell proliferation and induces apoptosis by inhibiting ErbB3 signaling in H1299 lung cancer cells. TK or CuD could be useful as a compound to treat lung cancer. Additionally, targeting ErbB3 may also be useful for treating lung cancer.

中文翻译：

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Trichosanthes kirilowi​​i与顺铂和培美曲塞联合使用的协同抗癌作用通过调节ErbB3增强H1299非小细胞肺癌细胞的凋亡

背景

肺癌是全球最常见的恶性肿瘤之一。为了治疗肺癌，开发并测试了多种抗癌药物，但由于耐药性而失败。在本研究中，我们测试了草药（例如TK和CuD）作为抗癌药物，以减少副作用和耐药性。

方法

通过MTT测定法测量细胞活力。细胞周期停滞的分析通过流式细胞仪进行。通过膜联蛋白V-FITC / PI测定法测量了葫芦素D诱导的细胞凋亡。我们进行了RTK试剂盒分析。通过蛋白质印迹分析测量p-ErbB3，p-STAT3，p-NF-κB和半胱氨酸蛋白酶的水平。通过免疫细胞化学测量ErbB3的核染色。STAT3和NF-κB的转录活性通过STAT3和NF-κB荧光素酶报告基因检测。

结果

我们在人NSCLC肿瘤细胞的原代培养中发现了TK与CDDP和PXD的协同作用。CDDP / PXD和TK或CuD的组合抑制了H1299细胞的增殖。CDDP / PXD和TK或CuD的组合可诱导H1299细胞中的亚G1和G2 / M细胞周期停滞。CDDP / PXD和TK或CuD的组合可诱导H1299细胞凋亡，调节凋亡分子，引起形态变化并抑制集落形成。我们发现，TK抑制p-ErbB3的表达和信号传导。CDDP / PXD和TK或CuD的组合可抑制H1299细胞中的p-AKT，p-Erk和p-JNK信号传导，并抑制Stat3和NF-κB转录活性。更重要的是，CDDP / PXD和TK或CuD的组合抑制了H1299细胞中的p-ErbB3和下游分子。CDDP / PXD和TK或CuD的组合抑制ErbB2 / ErbB3二聚化。我们的结果清楚地表明，CDDP / PXD与TK或CuD的协同作用可通过抑制ErbB3信号传导来抑制细胞生长并诱导细胞凋亡。

结论

CDDP / PXD和TK或CuD的组合可通过抑制H1299肺癌细胞中的ErbB3信号传导来降低细胞增殖并诱导凋亡。TK或CuD可用作治疗肺癌的化合物。另外，靶向ErbB3也可用于治疗肺癌。