Helicobacter pylori: Molecular detection of vacA gene and vacuolating activity in human gastric adenocarcinoma cells

Alfizah Hanafiah, Noraziah Mohamad Zin, M. Y. Chao, Md. Mostafizur Rahman, M. Ramelah

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Background and Aims: Helicobacter pylori strains secrete a vacuolating cytotoxin (VacA), plays an important role for the development of peptic ulcer disease and gastro-duodenal diseases. vacA gene is responsible to regulate the activity of the vacuolating cytotoxin. The objective of this study was molecular detection of vacA gene and observes the vacuolating activity on human gastric adenocarcinoma (AGS) cells. Materials and Methods: H. pylori vacA gene was determined by polymerase chain reaction. Vacuolation activity of VacA toxin in broth culture filtrates was assessed in AGS cells and quantified by neutral uptake assay. Different concentration dosages of VacA and incubation time were used in the measurement of the vacuolating activity on AGS cells. Results: The results showed that VacA toxin could stimulate vacuolating activity on AGS cells with minimum concentration 1.0 μg/ml from both of s1m1 and s1m2 alleles (vacA gene). The toxin produced optimal reaction at 5.0 μg/ml with significant differences observed between the alleles. The results also showed that both alleles commenced the vacuolating activity at the minimum of 3 hr incubation time, and the activity showed in time-dependent manner. Conclusion: Optimal concentration of VacA toxin (s1m1 allele) causes more interaction with AGS cell producing more vacuolating activities. Time-dependent vaculation of both alleles might allow H. pylori for persistent infection without rapid destruction of gastric cells might promote gastro-duodenal diseases. The study provided us better understanding of the pathogenesis of the diseases associated with H. pylori infection which is an emerging problem in developing countries.

Original languageEnglish
Pages (from-to)301-305
Number of pages5
JournalClinica Terapeutica
Volume164
Issue number4
DOIs
Publication statusPublished - 2013

Fingerprint

Human Activities
Helicobacter pylori
Stomach
Adenocarcinoma
Alleles
Duodenal Diseases
Genes
Cytotoxins
Helicobacter Infections
Peptic Ulcer
Developing Countries
Polymerase Chain Reaction
Infection

Keywords

  • Adenocarcinoma cells(AGS)
  • Helicobacter pylori
  • pathogenesis
  • Polymerase chain reaction(PCR)
  • vaca gene
  • Vacuolating cytotoxin

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Helicobacter pylori : Molecular detection of vacA gene and vacuolating activity in human gastric adenocarcinoma cells. / Hanafiah, Alfizah; Mohamad Zin, Noraziah; Chao, M. Y.; Rahman, Md. Mostafizur; Ramelah, M.

In: Clinica Terapeutica, Vol. 164, No. 4, 2013, p. 301-305.

Research output: Contribution to journalArticle

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N2 - Background and Aims: Helicobacter pylori strains secrete a vacuolating cytotoxin (VacA), plays an important role for the development of peptic ulcer disease and gastro-duodenal diseases. vacA gene is responsible to regulate the activity of the vacuolating cytotoxin. The objective of this study was molecular detection of vacA gene and observes the vacuolating activity on human gastric adenocarcinoma (AGS) cells. Materials and Methods: H. pylori vacA gene was determined by polymerase chain reaction. Vacuolation activity of VacA toxin in broth culture filtrates was assessed in AGS cells and quantified by neutral uptake assay. Different concentration dosages of VacA and incubation time were used in the measurement of the vacuolating activity on AGS cells. Results: The results showed that VacA toxin could stimulate vacuolating activity on AGS cells with minimum concentration 1.0 μg/ml from both of s1m1 and s1m2 alleles (vacA gene). The toxin produced optimal reaction at 5.0 μg/ml with significant differences observed between the alleles. The results also showed that both alleles commenced the vacuolating activity at the minimum of 3 hr incubation time, and the activity showed in time-dependent manner. Conclusion: Optimal concentration of VacA toxin (s1m1 allele) causes more interaction with AGS cell producing more vacuolating activities. Time-dependent vaculation of both alleles might allow H. pylori for persistent infection without rapid destruction of gastric cells might promote gastro-duodenal diseases. The study provided us better understanding of the pathogenesis of the diseases associated with H. pylori infection which is an emerging problem in developing countries.

AB - Background and Aims: Helicobacter pylori strains secrete a vacuolating cytotoxin (VacA), plays an important role for the development of peptic ulcer disease and gastro-duodenal diseases. vacA gene is responsible to regulate the activity of the vacuolating cytotoxin. The objective of this study was molecular detection of vacA gene and observes the vacuolating activity on human gastric adenocarcinoma (AGS) cells. Materials and Methods: H. pylori vacA gene was determined by polymerase chain reaction. Vacuolation activity of VacA toxin in broth culture filtrates was assessed in AGS cells and quantified by neutral uptake assay. Different concentration dosages of VacA and incubation time were used in the measurement of the vacuolating activity on AGS cells. Results: The results showed that VacA toxin could stimulate vacuolating activity on AGS cells with minimum concentration 1.0 μg/ml from both of s1m1 and s1m2 alleles (vacA gene). The toxin produced optimal reaction at 5.0 μg/ml with significant differences observed between the alleles. The results also showed that both alleles commenced the vacuolating activity at the minimum of 3 hr incubation time, and the activity showed in time-dependent manner. Conclusion: Optimal concentration of VacA toxin (s1m1 allele) causes more interaction with AGS cell producing more vacuolating activities. Time-dependent vaculation of both alleles might allow H. pylori for persistent infection without rapid destruction of gastric cells might promote gastro-duodenal diseases. The study provided us better understanding of the pathogenesis of the diseases associated with H. pylori infection which is an emerging problem in developing countries.

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