Alantolactone exerts anti-proliferative and apoptotic effects on BGC823 and SGC7901 cells via activation of p38MAPK and inhibition of NF-κB signaling pathway

Xie Ningsheng,Luo Qingtian,Gu Qiuping,Xie Junfeng,Zhu Fangqin Fangqin,Liu Youshun,Tang Jianhua,Wei Yingfeng

doi:10.4314/tjpr.v18i3.3

Abstract

Purpose: To investigate the anti-proliferative and apoptotic influences of alantolactone on gastric carcinoma (GC) cell lines, and the mechanism(s) involved. Methods: Human gastric cancer cell line (BGC823) and gastric adenocarcinoma lymph node metastasis cell line (SGC7901) were maintained in Ham’s F12 medium supplemented with 10 % heatinactivated fetal bovine serum (FBS). In each group of cancer cell line, 5 groups of cells were used: control and four alantolactone groups which were treated with increasing concentrations of alantolactone (5 - 30 μM) for varying periods. Proliferation was determined using MTT assay, while realtime quantitative polymerase chain reaction (qRT-PCR) was used to assay the expressions of apoptosis- and metastasis-related genes. The expressions of p38MAPK and nuclear transcription factor-κB (NF-κB) in BGC823 and SGC7901 cells were measured with Western blotting. Results: Phosphorylated protein (p-p38 protein) expression was significantly higher in both groups of GC cells, relative to control (p < 0.05). The expressions of NF-κB in plasma protein were markedly higher in both groups of GC cells than in control group, but the corresponding expressions in nuclear protein were significantly lower in both groups of GC cells, relative to control (p < 0.05). Conclusion: Alantolactone exerts anti-proliferative and apoptotic effects on BGC823 and SGC7901 cells via mechanisms involving activation of the p38MAPK, and inhibition of the NF-κB signaling pathways. Thus, alantolactone may be a new and effective anti-gastric cancer drug.

Highlights

Gastric cancer (GC), a malignant tumor of the gastric mucosa is characterized by high incidence and mortality
The expressions of p38MAPK), nuclear transcription factor-κB (NF-κB) in BGC823 and SGC7901 cells treated with alantolactone (20 μM, 24 h) were measured using Western blotting
The proliferations of BGC823 and SGC7901 cells were significantly inhibited in a time- and concentration-dependent manner by alantolactone, relative to control group (p < 0.05)

Summary

INTRODUCTION

Gastric cancer (GC), a malignant tumor of the gastric mucosa is characterized by high incidence and mortality. The cells (1 × 105 cells/mL) were seeded into 96-well plates and pretreated with varied concentrations of alantolactone (5 - 30 μM) This was followed by the addition of 20 mL of 0.5 % MTT solution within 4 h, after which the culture medium was changed. Biotechnology Co., Ltd. Phosphate buffered saline (PBS) was purchased from SGM Biotech and RIPA protein lysis kit was obtained from. The treated and control cells were lysed using radioimmunoprecipitation assay (RIPA) buffer. Their total RNAs were extracted using Trizol reagent, and reverse-transcribed to cDNAs, which were quantified using qRT-PCR. The expressions of p38MAPK), nuclear transcription factor-κB (NF-κB) in BGC823 and SGC7901 cells treated with alantolactone (20 μM, 24 h) were measured using Western blotting.

RESULTS

DISCUSSION

CONCLUSION

Conflict of Interest