title: Anti-proliferative effect of RCE-4 from Reineckia carnea on human cervical cancer HeLa cells by inhibiting the PI3K/Akt/mTOR signaling pathway and NF-κB activation.
abstract: Cervical cancer is the second leading cause of cancer deaths in women worldwide. In recent years, the studies find that inflammation is a critical component of tumor progression, and the ideal therapeutic methods should be aimed at the inflammation reaction triggers. (1β,3β,5β,25S)-spirostan-1,3-diol1-[α-L-rhamnopyranosyl-(1 → 2)-β-D-xylopyranoside] (RCE-4) was the main active composition of Reineckia carnea (Andr.) Kunth. It significantly induced apoptosis in cervical cancer Caski cells through the mitochondrial pathway in our previous studies; however, its underlying mechanism remains poorly understood. This study aimed to further evaluate the effect of RCE-4 on human cervical cancer HeLa cells. Based on this observation, we investigated the anti-cervical cancer effect of RCE-4 by modulating phosphatidylinositol 3-kinase/protein kinase-B/mammalian target of rapamycin (PI3K/Akt/mTOR) signaling pathway, nuclear factor-kappa B (NF-κB) activation, and inflammation-related key factors in HeLa cells. The results indicated that the HeLa cell was the most sensitive with an IC50 of 7.01 μM; RCE-4 significantly promoted the release of cellular lactate dehydrogenase (LDH); increased DNA fragmentation and apoptosis; reduced PI3K, Akt, mTOR, and NF-κBp65 phosphorylation levels; increased the Bax and cleaved poly (ADP-ribose) polymerase (PARP) protein levels; suppressed Bcl-2 protein expression; elevated the Bax/Bcl-2 expression ratio; and decreased the interleukin-1 beta (IL-1β) and interleukin-6 (IL-6) mRNA expressions in HeLa cells in a concentration-dependent manner. These findings suggest that RCE-4 exerted beneficially anti-cervical cancer effect on HeLa cells, mainly inhibiting PI3K/Akt/mTOR signaling pathway phosphorylation and NF-κB activation, promoting HeLa cell apoptosis. Graphical abstract Anti-tumor effect of RCE-4 on HeLa cells.
authors.affiliation: Hubei Key Laboratory of Natural Products Research and Development, China Three Gorges University, 8, University Avenue, 443002, Yichang, Hubei, China.
authors.forename: Caihong
authors.initials: C
authors.lastname: Bai
authors.affiliation: Hubei Key Laboratory of Natural Products Research and Development, China Three Gorges University, 8, University Avenue, 443002, Yichang, Hubei, China.
authors.forename: Xiaojiao
authors.initials: X
authors.lastname: Yang
authors.affiliation: Hubei Key Laboratory of Natural Products Research and Development, China Three Gorges University, 8, University Avenue, 443002, Yichang, Hubei, China.
authors.forename: Kun
authors.initials: K
authors.lastname: Zou
authors.affiliation: Hubei Key Laboratory of Natural Products Research and Development, China Three Gorges University, 8, University Avenue, 443002, Yichang, Hubei, China. hehaibo252219@163.com.|Hubei Key Laboratory of Tumor Microenvironment and Immunotherapy, China Three Gorges University, Yichang, Hubei, China. hehaibo252219@163.com.
authors.forename: Haibo
authors.initials: H
authors.lastname: He
authors.affiliation: Hubei Key Laboratory of Natural Products Research and Development, China Three Gorges University, 8, University Avenue, 443002, Yichang, Hubei, China.
authors.forename: Junzhi
authors.initials: J
authors.lastname: Wang
authors.affiliation: Hubei Key Laboratory of Natural Products Research and Development, China Three Gorges University, 8, University Avenue, 443002, Yichang, Hubei, China.
authors.forename: Huilin
authors.initials: H
authors.lastname: Qin
authors.affiliation: Hubei Key Laboratory of Natural Products Research and Development, China Three Gorges University, 8, University Avenue, 443002, Yichang, Hubei, China.
authors.forename: Xiaoqin
authors.initials: X
authors.lastname: Yu
authors.affiliation: Hubei Key Laboratory of Natural Products Research and Development, China Three Gorges University, 8, University Avenue, 443002, Yichang, Hubei, China.
authors.forename: Chengxiong
authors.initials: C
authors.lastname: Liu
authors.affiliation: Hubei Key Laboratory of Natural Products Research and Development, China Three Gorges University, 8, University Avenue, 443002, Yichang, Hubei, China.
authors.forename: Juyan
authors.initials: J
authors.lastname: Zheng
authors.affiliation: Hubei Key Laboratory of Natural Products Research and Development, China Three Gorges University, 8, University Avenue, 443002, Yichang, Hubei, China.
authors.forename: Fan
authors.initials: F
authors.lastname: Cheng
authors.affiliation: Hubei Key Laboratory of Natural Products Research and Development, China Three Gorges University, 8, University Avenue, 443002, Yichang, Hubei, China.
authors.forename: Jianfeng
authors.initials: J
authors.lastname: Chen
mesh_terms: D000972:Antineoplastic Agents, Phytogenic / Q000302:isolation & purification / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D016207:Cytokines / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D005260:Female; D006367:HeLa Cells; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism; D020128:Inhibitory Concentration 50; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D019668:Liliaceae / Q000737:chemistry; D061986:MCF-7 Cells; D016328:NF-kappa B / Q000378:metabolism*; D058539:Phosphatidylinositol 3-Kinase / Q000378:metabolism*; D010766:Phosphorylation; D008517:Phytotherapy; D010946:Plants, Medicinal; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D012503:Saponins / Q000302:isolation & purification / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects; D013150:Spirostans / Q000302:isolation & purification / Q000494:pharmacology*; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism*; D002583:Uterine Cervical Neoplasms / Q000188:drug therapy* / Q000201:enzymology / Q000473:pathology
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D051017:Apoptosis Regulatory Proteins; D016207:Cytokines; D018836:Inflammation Mediators; D016328:NF-kappa B; C580846:RCE-4 compound; D012503:Saponins; D013150:Spirostans; D007770:L-Lactate Dehydrogenase; C546842:MTOR protein, human; D058539:Phosphatidylinositol 3-Kinase; D051057:Proto-Oncogene Proteins c-akt; D058570:TOR Serine-Threonine Kinases
keywords: Apoptosis; HeLa cells; Inflammatory factor; NF-κB; PI3K/Akt/mTOR