Objective: To analyze highly expressed genes in Tetrastigma hemsleyanum roots and the in vitro antitumor effects of extracts from different parts of this plant. Methods: The expression profiles of the roots, stems, and leaves of T hemsleyanum were analyzed via high-throughput sequencing technology with the Illumina Genome Analyzer platform. Then, gene ontology (GO) functional and pathway enrichment analyses of the significantly differentially expressed genes were performed. The inhibition rates of the extracts from different plant parts were determined through the CCK-8 method. Results: Genes related to catalytic activity and cytochrome P450 were upregulated in roots compared with leaves. The genes that we screened that were expressed at high levels in the roots may play an important role in the accumulation of active ingredients. GO functional enrichment analysis revealed that there were substantial differences in genes related to metabolic processes in roots compared with those in stems, and KEGG analysis revealed significant differences in genes related to the zeatin biosynthesis and diterpenoid biosynthesis pathways. Moreover, the expression of genes related to catalytic activity, ion binding, and hydrolase activity differed significantly between the roots and leaves, and KEGG analysis revealed significant differences in the expression of genes related to terpenoid biosynthesis and flavonoid biosynthesis pathways. Quantitative PCR indicated that Thhg was highly expressed in roots. The total triterpene content was also high in the tissue. The CCK-8 assays showed that the root ethanol extract had the highest antitumor activity. Conclusion: The genes we screened that were expressed at high levels in the roots may play an important role in the accumulation of active ingredients. Importantly, stem and leaf ethanol extracts also showed certain antitumor effects on human HCC Huh-7 and Hep3B cells in vitro and may also have medicinal value.

中文翻译：

---

四柱差异表达基因及其抗肿瘤活性研究

目的：分析四柱根高表达基因及不同部位提取物的体外抗肿瘤作用。方法：利用Illumina基因组分析仪平台，通过高通量测序技术对Themsleyanum根、茎、叶的表达谱进行分析。然后，对显着差异表达的基因进行基因本体（GO）功能和通路富集分析。通过CCK-8法测定植物不同部位提取物的抑制率。结果：与叶相比，根中与催化活性和细胞色素P450相关的基因上调。我们筛选出的在根部高水平表达的基因可能在活性成分的积累中发挥重要作用。 GO功能富集分析显示根部与茎部代谢过程相关基因存在显着差异，KEGG分析显示玉米素生物合成和二萜生物合成途径相关基因存在显着差异。此外，根和叶中催化活性、离子结合和水解酶活性相关基因的表达存在显着差异，KEGG分析显示萜类生物合成和类黄酮生物合成途径相关基因的表达存在显着差异。定量PCR表明Thhg在根中高表达。组织中总三萜含量也很高。 CCK-8测定表明根乙醇提取物具有最高的抗肿瘤活性。结论：我们筛选到的根部高表达基因可能在活性成分的积累中发挥重要作用。重要的是，茎叶乙醇提取物在体外对人肝癌Huh-7和Hep3B细胞也表现出一定的抗肿瘤作用，也可能具有药用价值。