Application of Electric Cell-Substrate Impedance Sensing to Investigate the Cytotoxic Effects of Andrographolide on U-87 MG Glioblastoma Cell Migration and Apoptosis

Abstract

Glioblastoma multiforme (GBM) is the most common and aggressive primary brain tumor in adults. In recent studies, the efficacy of suberoylanilide hydroxamic acid (SAHA) has been investigated for GBM. We explored the effects of two exploratory compounds, the histone deacetylase SAHA and the natural product andrographolide, on Uppsala 87 Malignant Glioma (U-87 MG) cell migration and viability in comparison with the clinically used therapeutic agent temozolomide (TMZ). We used the electric cell–substrate impedance sensing (ECIS) system to monitor the migration of U-87 MG cells after treatment with various concentrations of these compounds. Moreover, we used the Alamar blue assay and western blotting to observe the concentration-dependent changes in the viability and apoptosis of U-87 MG cells. Our results demonstrated that both SAHA and andrographolide (10–300 μM) significantly inhibited GBM cell migration in a concentration-dependent manner, and 10 μM SAHA and 56 μM andrographolide demonstrated remarkable inhibitory effects on U-87 MG migration. Western blotting indicated that compared with TMZ, both SAHA and andrographolide induced higher expression levels of apoptosis-related proteins, such as caspase-3, BAX, and PARP in U-87 MG cells. Furthermore, all three drugs downregulated the expression of the antiapoptotic protein Bcl-2. In conclusion, SAHA and andrographolide showed exceptional results in inhibiting cell migration and motility. The ECIS wound healing assay is a powerful technique to identify and screen potential therapeutic agents that can inhibit cancer cell migration.

Application of ECIS to Assess FCCP-Induced Changes of MSC Micromotion and Wound Healing Migration

“…We have previously reported the use of ECIS wound healing assay to investigate cytotoxic effects of andrographolide on U-87 MG glioblastoma cell migration [28]. In the present study, we have used ECIS to assess hMSC wound-healing migration in response to low levels of FCCP and demonstrated the effect of FCCP-induced mitochondrial dysfunction on cell migration rate in a dose-dependent manner.…”

“…Fluctuations in impedance measured with this device have experimentally been related to cell micromotion [24,25,26,27]. Recently, we have applied ECIS wound-healing assay to investigate the cytotoxic effects of andrographolide on U-87 MG glioblastoma cell migration [28]. In this study, we used ECIS to sensitively detect the effect of different low levels of FCCP on hMSC micromotion and wound healing migration.…”

Andrographolide: A review of its pharmacology, pharmacokinetics, toxicity and clinical trials and pharmaceutical researches

“…In malignant glioma cell U‐87 MG, andrographolide evidently inhibited glioblastoma multiforme (GBM) cell migration in a concentration‐dependent manner. Equally important, it accelerated higher expression of apoptosis‐related proteins including caspase‐3, BAX, and PARP (Chiu, Batsaikhan, Huang, & Wang, 2019). In addition, researchers relies on the truth that andrographolide forcefully suppressed proliferation via inducing G 2 /M cell‐cycle arrest in human malignant melanoma A375 and C8161 cell lines (Liu & Chu, 2018).…”

Biological properties of andrographolide, an active ingredient of Andrographis Paniculata: a narrative review

“…Inhibition to migration was documented in the glioblastoma cell model where motility was impeded by downregulation of the ERK1/2-mediated matrix metalloproteinase-2 (MMP-2) expression (39,40). This is important as an anti-metastatic effect of AG described for Ras-transformed cells, like the glioblastoma (41).…”