The Novel Nature Microtubule Inhibitor Ivalin Induces G2/M Arrest and Apoptosis in Human Hepatocellular Carcinoma SMMC-7721 Cells In Vitro

Abstract

Background and Objectives: Microtubules are an attractive target for cancer chemotherapy. Previously, we reported that Ivalin exhibited excellent anti-migration and anti-invasion activities in human breast cancer cells. Here, we examined the microtubule inhibition effect of Ivalin in human hepatocellular carcinoma SMMC-7721 cells. Materials and Methods: We used the 3-(4,5-dimethylthiazol)-2,5-diphenyltetrazolium bromide (MTT) assay to evaluate the cell proliferation effect of Ivalin and flow cytometry analysis to detect the apoptotic and cell cycle arrest effects of Ivalin. Immunofluorescence staining was used to measure the effect of Ivalin on the cytoskeleton network, and Western blotting was used to detect the expression levels of Bax, Bcl-2, Cdc2, phosphor-Cdc2, Cdc25A, Cyclin B1, and tubulin. Results: Ivalin induced cell cycle G2/M arrest and subsequent triggered apoptosis in human hepatocellular carcinoma SMMC-7721 cells. Furthermore, microtubules were shown to be involved in Ivalin-meditated apoptosis. In this connection, Ivalin treatment suppressed cellular microtubule network formation by regulating microtubule depolymerization. Moreover, Western blotting revealed Cdc25A and Cyclin B1 were upregulated in Ivalin-meditated cell cycle arrest. Subsequently, the induction of Bax (a proapoptotic protein) and reduction of Bcl-2 (an anti-apoptotic protein) expression were observed in Ivalin-treated SMMC-7721 cells. Conclusion: Ivalin induced microtubule depolymerization, then blocked cells in mitotic phase, and eventually resulted in apoptosis in SMMC-7721 cells. Collectively, these data indicate that Ivalin, acting as a novel inhibitor of microtubules, could be considered as a promising lead in anticancer drug development.

Ivalin Induces Mitochondria-Mediated Apoptosis Associated with the NF-κB Activation in Human Hepatocellular Carcinoma SMMC-7721 Cells

“…Our previous studies confirmed that Ivalin (Figure 1) was significantly cytotoxic to SMMC-7721 cells (IC50: 4.34 ± 0.10) with a lower effect toward the normal cell line HL7702 (IC50: 25.86 ± 0.87) [13]. In response to characterizing the cell growth inhibition effect of Ivalin, we monitored morphological changes in SMMC-7721 cells after 24 h of treatment.…”

“…In our previous work, we reported the ideal microtubule depolymerization activities of Ivalin in SMMC-7721 cells [13]. In this study, we found that Ivalin treatment may lead to obviously apoptotic features including apoptotic body formation and nuclear condensation in the same cells.…”

“…The apoptosis rate of SMMC-7721 cells after Ivalin 24 h treatment was evaluated by Annexin V-FITC and Propidium Iodide (PI) double staining, according to our previously described study [13].…”

“…Ivalin (Figure 1), another compound with an eudesmane framework, was abstracted from the traditional herb Carpesium divaricatum [12]. Our previous works demonstrated that Ivalin can serve as a novel microtubule inhibitor by depolymerizing microtubules and resulted in cell proliferation inhibition in hepatocellular carcinoma SMMC-7721 cells [13]. Here, we report that Ivalin enforced the procedure of apoptosis in the same cells by triggering reactive oxygen species (ROS) generation and the participation of the mitochondria pathway.…”

Identification of hepatocellular carcinoma prognostic markers based on 10-immune gene signature

“…Studies have shown that Aberrant KIF20A expression might independently predict poor overall survival and recurrence-free survival of hepatocellular carcinoma [27]. CCNB1 is a cycle-like protein that regulates cell cycle G2/M, and its expression imbalance is one of the causes of malignant tumor proliferation, including HCC [28]. Researches show that disease-free survival in the high PLOD2 expression group of HCC patients was significantly shorter when compared with the low-expression group [29].…”