CC12 Induces Apoptotic Cell Death and Cell Cycle Arrest in Human Glioblastoma Cell Lines and Mouse Xenograft Model

Abstract

Among central nervous system tumors, glioblastoma (GBM) is the most common and the most malignant type. Even under current standard treatments, the overall survival rate is still low and the recurrence rate is high. Therefore, developing novel and effective therapy is urgently needed. CC12, a synthesized small molecule, was evaluated for the potential anti-GBM effects in two GBM cell lines, U87MG and U118MG. The observations of cell morphology, MTT assay, flow cytometry-based apoptosis after CC12 treatment, were conducted. Western blot was performed for the investigation of the apoptotic mechanism. Positron emission tomography scan analysis and bioluminescent imaging assay using a mouse xenograft model were performed for the effect of CC12 in vivo. After treated by 10 μM CC12 for 24 h, both U118MG and U87MG cells showed tumor cell death. MTT assay results showed that the survival rates decreased when the CC12 concentrations or the treatment periods increased. Ki-67 expression and flow cytometry results indicated that the proliferation was inhibited in GBM cells, and G1 phase arrest was shown. The results of 7-AAD, Br-dUTP, and JC-1 staining all showed the apoptosis of GBM cells after CC12 treatment. Increased γH2AX, caspase-3, and poly (ADP-ribose) polymerase (PARP) levels meant the DNA damage, and increased Bcl2 family proteins after CC12 treatment indicated the intrinsic apoptotic pathway was involved in CC12 induced apoptosis. Furthermore, CC12 can induce the decrease of tumor prognostic marker DcR3. In vivo experiment results showed the effect of CC12 on tumor size reduction of CC12. In addition, the ability to cross the brain–blood barrier of CC12 was also confirmed. CC12 may have anti-tumor ability through the regulation of cell cycle and apoptosis in vitro and in vivo.

Microfluidic model of the alternative vasculature in neuroblastoma

“…Cancer cells cultured in 2D often have little resistance to drugs [22][23][24]. For example, cancer cells are most sensitive to pro-apoptotic treatments when culture in plastic dishes may not be translatable to the clinic [25]. Importantly, 2D cancer models lack cell heterogeneity observed in clinical samples in patients.…”

In Vitro and Computational Studies of Perezone and Perezone Angelate as Potential Anti-Glioblastoma Multiforme Agents

“…The apoptotic cell ratio was measured by Annexin V/7AAD staining followed by flow cytometry analysis. Simultaneous staining of cells with Annexin V/7-AAD allows the discrimination of intact cells (Annexin V-/7-AAD-), early apoptotic (Annexin V+/7-AAD —), and late apoptotic or necrotic cells (Annexin V+/7-AAD +) [ 46 ]. Briefly, U373 cells (1 mL containing 10 6 per well) were seeded into 24-well microplates and incubated by 24 h. Then, 1 mL of solutions with increasing concentrations of perezone and perezone angelate diluted in fresh medium containing 2% ethanol were added to the respective wells, obtaining a final concentration of 6.25 and 50 μM ( n = 3) and incubated for an additional 48 h (final concentration of ethanol 1%), employing cells without treatment as control.…”

Inhibition of Proliferation and Induction of Apoptosis in Prostatic Carcinoma DU145 Cells by Polysaccharides from Yunnan Rosa roxburghii Tratt

“…Studies showed that gypenoside LI exhibits the ability to inhibit the proliferation and migration of human breast cancer cells, induce apoptosis, and arrest the cell cycle at the G0/G1 phase by modulating E2F1 [14]. Fann and colleagues found that treatment with anthraquinone (CC12) resulted in reduced Bcl2 protein expression in two glioblastoma cell lines, 87MG and U118MG, suggesting that CC12 induces apoptosis in tumor cells and arrests cells in the G1 phase [15].…”