Abstract：
Lung cancer poses a significant threat globally, especially in China. This puts higher demands on the treatment methods and drugs for lung cancer. Natural plants provide valuable resources for the development of anti-cancer drugs. Hederagenin (Hed) is a triterpenoid compound extracted from ivy leaves and has anti-tumor activity against multifarious cancers, including lung cancer. However, the regulatory mechanism of Hed in lung cancer remains unclear. In this study, we used Hed to treat lung cancer cells, and observed the effect of Hed on cell proliferation (including CCK-8 and colony formation experiments), apoptosis (including flow cytometry and apoptosis gene detection (BAX and Bcl-2)). The results showed that Hed induced lung cancer cell death (inhibiting proliferation and promoting apoptosis). Next, we performed bioinformatics analysis of the expression profile GSE186218 and found that Hed treatment significantly increased the expression of CHAC1 gene. CHAC1 is a ferroptosis-inducing gene. RT-qPCR detection of lung cancer clinical tissues and related cell lines also showed that CHAC1 was lowly expressed in lung cancer. Therefore, we knocked down and overexpressed CHAC1 in lung cancer cells, respectively. Subsequently, cell phenotype experiments showed that down-regulating CHAC1 expression inhibited lung cancer cell death (promoting proliferation and inhibiting apoptosis); on the contrary, up-regulating CHAC1 expression promoted lung cancer cell death. To further verify that Hed exerts anti-tumor effects in lung cancer by promoting CHAC1 expression, we performed functional rescue experiments. The results showed that down-regulating CHAC1 expression reversed the promoting effect of Hed on lung cancer cell death. Mechanistically, in vitro and in vivo experiments jointly demonstrated that Hed exerts anti-cancer effects by promoting CHAC1-induced ferroptosis. In summary, our study further enriches the regulatory mechanism of Hed in lung cancer.
摘要：
肺癌在全球范围内构成重大威胁，尤其是在中国。这对肺癌的治疗方法和药物提出了更高的要求。天然植物为抗癌药物的开发提供了宝贵的资源。Hederagenin（Hed）是从常春藤叶中提取的三萜化合物，对多种癌症具有抗肿瘤活性，包括肺癌.然而,Hed在肺癌中的调控机制尚不清楚。在这项研究中,我们用赫德治疗肺癌细胞，并观察了Hed对细胞增殖的影响（包括CCK-8和集落形成实验），细胞凋亡(包括流式细胞术和凋亡基因检测(BAX和Bcl-2))。结果表明，Hed诱导肺癌细胞死亡（抑制增殖，促进凋亡）。接下来,我们对GSE186218的表达谱进行了生物信息学分析，发现Hed处理显著增加了CHAC1基因的表达。CHAC1是铁凋亡诱导基因。RT-qPCR检测肺癌临床组织和相关细胞系也显示CHAC1在肺癌中低表达。因此,我们在肺癌细胞中敲低并过表达CHAC1，分别。随后，细胞表型实验表明下调CHAC1表达抑制肺癌细胞死亡(促进增殖和抑制凋亡),上调CHAC1表达促进肺癌细胞死亡。为了进一步验证Hed通过促进CHAC1的表达在肺癌中发挥抗肿瘤作用,我们进行了功能性救援实验。结果表明，下调CHAC1的表达可逆转Hed对肺癌细胞死亡的促进作用。机械上,体外和体内实验共同证明Hed通过促进CHAC1诱导的铁凋亡发挥抗癌作用。总之,本研究进一步丰富了Hed在肺癌中的调控机制。
