Effects of Andrographolide on Intracellular pH Regulation, Cellular Migration, and Apoptosis in Human Cervical Cancer Cells (Running Tittle: Effects of Andrographolide on pH Regulators and Apoptosis in Cervical Cancer)

Abstract

Cancer cells have been characterized with alkaline intracellular pH (pHi) values (≥7.2) to enable cancer proliferation, migration, and progression. The aim of the present study was to explore the concentration-dependent effects of Andrographolide, an active diterpenoid compound of herb Andrographis paniculata, on Na+/H+ exchanger isoform 1 (NHE1), cellular migration and apoptosis in human cervical cancer cells (HeLa). The pHi was detected by microspectrofluorometry method, and intracellular acidification was induced by NH4Cl prepulse technique. Viability and protein expression were determined by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and Western blot, respectively. Human normal endocervical cells (End1), ectocervical cells (Ect1), and HeLa were bought commercially. The resting pHi value of HeLa (≈7.47) was significantly higher than that of End1 and Ect1 (≈7.30), and shifted from alkaline to acidic following acid/base impacts. In HEPES (4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid | N-(2-Hydroxyethyl)piperazine-N′-(2-ethanesulfonic acid) -buffered superfusate, NHE1 and V-ATPase co-existed functionally for acid extrusion in HeLa, while only NHE1 existed functionally in End/Ect1. Andrographolide (3–1000 μM) concentration-dependently inhibited NHE1 activity. Cell-migration and expressions of NHE1, V-ATPase, PARP (poly-ADP-ribose-polymerase), pro-Caspase-3, and Bcl-2 were significantly reduced by pretreating with Andrographolide (≥100 μM) for 24–48 h in HeLa. Andrographolide inhibited cell viability of End1-cells/Ect1 and HeLa (≥100 and ≥30 μM, respectively). The present findings implicate the promising clinical applications of Andrographolide on cervical cancer treatment.

Advances in research on the regulatory mechanism of NHE1 in tumors (Review)

“…An experiment by Chiang et al demonstrated that EGF is involved in the regulation of NHE1 expression via the PI3K signaling pathway in cervical cancer (91), and NHE1 can also interact with the actin-related protein ezrin to reconstitute the cytoskeleton and stimulate the migration and invasion of cervical cancer cells. It was also reported that Andrographolide regulates the expression of apoptotic proteins to induce cells apoptosis, and concentration-dependently decreases pHi by decreasing the activity of NHE1/V-ATPase and the expression of NHE1 in HeLa cells; thus, Andrographolide may represent a promising novel agent in the treatment of cervical cancer in the clinical setting (92). In addition, NHE has also been found to be involved in the regulation of uterine fluid pH (Fig.…”

Andrographis paniculata (Burm. f.) Wall. ex Nees: An Updated Review of Phytochemistry, Antimicrobial Pharmacology, and Clinical Safety and Efficacy

“…Modern science is focusing on validating the traditional claims of this plant through systemic investigations. Different extracts (i.e., acetone, chloroform, ethanol, hexane, methanolic or aqueous extract) and isolated pure metabolites from A. paniculata have been investigated for pharmacological properties, for example, antibacterial, antiviral, antifungal, antiparasitic, choleretic, hypocholesterolemia, anti-inflammatory, anti-hyperglycemic, hepatoprotective, anticancer, immunomodulatory, cardiovascular, antihyperlipidemic, emollient, anti-snake venom, anti-platelet aggregation, anti-fertility, carminative, and antipyretic properties at in vitro and in vivo conditions [41][42][43][44][45][46][47][48][49][50][51][52][53].…”

Advances in ameliorating inflammatory diseases and cancers by andrographolide: Pharmacokinetics, pharmacodynamics, and perspective

“…Beyond the above non‐solid tumors, According to Loh et al, andrographolide inhibited cell survival and migration in human cervical cancer cells. This study showed therapeutic effects in treating cervical cancer may occur in andrographolide 256 . According to increasing evidence, Andrographolide has been investigated as a leading component in cancer treatment.…”

“…This study showed therapeutic effects in treating cervical cancer may occur in andrographolide. 256 According to increasing evidence, Andrographolide has been investigated as a leading component in cancer treatment. There are still several obstacles to better promoting andrographolide and its analogs as cancer therapy agents.…”