Department of Life Science, Dongguk University-Seoul, Goyang, Republic of Korea.
Am J Chin Med. 2022;50(6):1703-1717. doi: 10.1142/S0192415X22500720. Epub 2022 Jul 4.
While a number of coding genes have explained the anticancer activity of ginsenoside Rh2, little is known about noncoding RNAs. This study was performed to elucidate the regulatory activity of long noncoding RNA (lncRNA) CFAP20DC-AS1, which is known to be downregulated by Rh2. MiR-3614-3p, which potentially binds CFAP20DC-AS1, was screened using the LncBase Predicted program, and the binding was verified by assaying the luciferase activity of a luciferase/lncRNA recombinant plasmid construct. The competitive endogenous RNA (ceRNA) relationship of the two RNAs was further validated by quantitative PCR after deregulation of each RNA using siRNA. The effect of miRNA and target genes on the MCF-7 cancer cell growth was determined by monitoring proliferation and apoptosis in the presence of Rh2 after deregulating the corresponding gene. The miRNA decreased the luciferase activity of the luciferase/CFAP20DC-AS1 fusion vector, confirming the binding. SiRNA-based deregulation of CFAP20DC-AS1 attenuated the expression of miR-3614-3p and . In contrast to CFAP20DC-AS1, miR-3614-3p was upregulated by Rh2, inhibiting proliferation but stimulating apoptosis of the MCF-7 cells. Target genes of miR-3614-3p, BBX and TNFAIP3, were downregulated by Rh2 and the miRNA but upregulated by the lncRNA. Rh2 inhibits CFAP20DC-AS1, which obscures the association of the lncRNA with miR-3614-3p, resulting in the suppression of oncogenic BBX and TNFAIP3. Taken together, the Rh2/CFAP20DC-AS1/miR-3614-3p/target gene axis contributes to the antiproliferation activity of Rh2 in cancer cells.
虽然许多编码基因已经解释了人参皂苷 Rh2 的抗癌活性，但对于非编码 RNA 知之甚少。本研究旨在阐明长链非编码 RNA (lncRNA) CFAP20DC-AS1 的调节活性，已知 Rh2 下调 CFAP20DC-AS1 的表达。使用 LncBase Predicted 程序筛选可能与 CFAP20DC-AS1 结合的 miR-3614-3p，并通过测定荧光素酶/lncRNA 重组质粒构建体的荧光素酶活性来验证结合。使用 siRNA 下调每种 RNA 后，通过定量 PCR 进一步验证了两种 RNA 的竞争内源性 RNA (ceRNA) 关系。在下调相应基因后，用 Rh2 监测 MCF-7 癌细胞生长中的增殖和凋亡，确定 miRNA 和靶基因对 MCF-7 癌细胞生长的影响。miRNA 降低了荧光素酶/CFAP20DC-AS1 融合载体的荧光素酶活性，证实了结合。基于 siRNA 的 CFAP20DC-AS1 下调减弱了 miR-3614-3p 的表达。与 CFAP20DC-AS1 相反，Rh2 上调 miR-3614-3p，抑制 MCF-7 细胞的增殖但刺激其凋亡。miR-3614-3p 的靶基因 BBX 和 TNFAIP3 被 Rh2 和 miRNA 下调，但被 lncRNA 上调。Rh2 抑制 CFAP20DC-AS1，从而掩盖了 lncRNA 与 miR-3614-3p 的关联，导致致癌基因 BBX 和 TNFAIP3 被抑制。总之，Rh2/CFAP20DC-AS1/miR-3614-3p/靶基因轴有助于 Rh2 在癌细胞中的抗增殖活性。
