Background:Our previous clinical study has shown that Chinese herbal medicine (CHM) Fuzheng Kang-Ai (FZKA) decoction is effective in treating advanced lung cancer patients through prolonging the drug resistance to Gefitinib (GFTN). Our basic study found that FZKA decoction could enhance the inhibition effect of GFTN in lung cancer by inactivating PI3K/Akt pathway. Moreover, our recent work showed that FZKA induced lung cancer cell apoptosis via STAT3/Bcl-2/Caspase-3 pathway. Thus in this study, we aim to elucidate how FZKA enhances the effect of GFTN in lung cancer from the perspective of cell apoptosis.
Methods:Cell proliferation and colony formation assay were performed to detect the cell growth inhibition. Flow cytometry and TUNEL assay were carried out to test the cell apoptosis. Mitochondrial membrane potential (MMP) assay was done to measure the alteration of MMP. Caspase-3/-9 activity assay was used to test the activity of caspase-3/-9. Western blot and qRT-PCR were done to detect the expression of STAT3 and Bcl-2 family as well as Caspase-3/-9 and Cyt-C at protein and mRNA levels, respectively. Transient transfection was performed to silence STAT3 using siSTAT3. Animal model was done to validate the molecular mechanisms in vivo and immunohistochemistry was done to detect the expression of Bax and Caspase-3.
Results:Firstly, our results showed that FZKA enhanced the inhibition effect of GFTN in lung cancer both in vitro and in vivo. Secondly, cell apoptosis was enhanced when treating lung cancer cells with both FZKA and GFTN, a process involving the mitochondria and the Bcl-2 family. And Bcl-2 family was involved in this process. Interestingly, STAT3 plays a critical role on mediating the above process. Last but not the least, the enhanced effect of cell apoptosis induction of GFTN by FZKA was validated in animal model.
Conclusion:Our findings conclude that Fuzheng Kang-Ai decoction enhances the effect of GFTN-induced cell apoptosis in lung cancer through the mitochondrial pathway, providing a novel molecular mechanism by which FZKA sensitizes to GFTN by delaying drug resistance in treating lung cancer patients.
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【Fuzheng Kang-Ai decoction enhances the effect of Gefitinib-induced cell apoptosis in lung cancer through mitochondrial pathway.].
【扶正康爱汤通过线粒体途径增强吉非替尼诱导的肺癌细胞凋亡的作用。】
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DOI：10.1186/s12935-020-01270-3文章类型：杂志文章
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背景：我们先前的临床研究表明，中药复方抗癌汤可通过延长对吉非替尼（GFTN）的耐药性来有效治疗晚期肺癌患者。我们的基础研究发现，FZKA汤可通过灭活PI3K / Akt途径来增强GFTN对肺癌的抑制作用。此外，我们最近的工作表明FZKA通过STAT3 / Bcl-2 / Caspase-3途径诱导肺癌细胞凋亡。因此，在这项研究中，我们旨在从细胞凋亡的角度阐明FZKA如何增强GFTN在肺癌中的作用。
方法：通过细胞增殖和集落形成实验检测细胞的生长抑制情况。进行流式细胞术和TUNEL法检测细胞凋亡。进行了线粒体膜电位（MMP）测定以测量MMP的变化。 Caspase-3 / -9活性测定用于测试caspase-3 / -9的活性。进行了蛋白质印迹和qRT-PCR来分别检测STAT3和Bcl-2家族以及Caspase-3 / -9和Cyt-C在蛋白质和mRNA水平上的表达。使用siSTAT3进行瞬时转染以使STAT3沉默。建立动物模型以验证体内分子机制，并进行免疫组织化学以检测Bax和Caspase-3的表达。
结果：首先，我们的结果表明，FZKA在体外和体内均增强了GFTN对肺癌的抑制作用。其次，当用FZKA和GFTN处理肺癌细胞时，细胞凋亡得到增强，这一过程涉及线粒体和Bcl-2家族。 Bcl-2家族参与了这一过程。有趣的是，STAT3在介导上述过程中起着至关重要的作用。最后但并非最不重要的一点是，在动物模型中验证了FZKA增强GFTN诱导细胞凋亡的作用。
结论：我们的研究结论表明，扶正抗癌汤通过线粒体途径增强了GFTN诱导的肺癌细胞凋亡的作用，为FZKA延缓耐药性对肺癌的耐药提供了新的分子机制。
方法：通过细胞增殖和集落形成实验检测细胞的生长抑制情况。进行流式细胞术和TUNEL法检测细胞凋亡。进行了线粒体膜电位（MMP）测定以测量MMP的变化。 Caspase-3 / -9活性测定用于测试caspase-3 / -9的活性。进行了蛋白质印迹和qRT-PCR来分别检测STAT3和Bcl-2家族以及Caspase-3 / -9和Cyt-C在蛋白质和mRNA水平上的表达。使用siSTAT3进行瞬时转染以使STAT3沉默。建立动物模型以验证体内分子机制，并进行免疫组织化学以检测Bax和Caspase-3的表达。
结果：首先，我们的结果表明，FZKA在体外和体内均增强了GFTN对肺癌的抑制作用。其次，当用FZKA和GFTN处理肺癌细胞时，细胞凋亡得到增强，这一过程涉及线粒体和Bcl-2家族。 Bcl-2家族参与了这一过程。有趣的是，STAT3在介导上述过程中起着至关重要的作用。最后但并非最不重要的一点是，在动物模型中验证了FZKA增强GFTN诱导细胞凋亡的作用。
结论：我们的研究结论表明，扶正抗癌汤通过线粒体途径增强了GFTN诱导的肺癌细胞凋亡的作用，为FZKA延缓耐药性对肺癌的耐药提供了新的分子机制。
