Gene expression in individual cells: analysis using global single cell reverse transcription polymerase chain reaction (GSC RT-PCR)

1999 ◽  
Vol 406 (2-4) ◽  
pp. 45-54 ◽  
Author(s):  
Leslie H Brail ◽  
Anne Jang ◽  
Filio Billia ◽  
Norman N Iscove ◽  
Henry J Klamut ◽  
...  
2020 ◽  
Vol 8 (A) ◽  
pp. 146-152
Author(s):  
Taufiq Qurrohman ◽  
Mohammad Basyuni ◽  
Poppy Anjelisa Zaitun Hasibuan

BACKGROUND: According to the Global Cancer Observatory in 2018, Asia was the first to note the incidence of colon cancer, which was 51.8% of cases of colon cancer which ranked the top three in the number of causes of death in the world. Cancer is a disease characterized by uncontrolled cell growth. Potential natural ingredient developed as chemotherapeutic agents includes from mangrove leaves. Studies reporting on the pharmacological activity of polyisoprenoid from mangrove species are still limited, therefore, it is essential to achieve the prospects, potential, and mechanisms polyisoprenoid in mangroves as a natural ingredient of pharmaceutical and medication. AIM: The aim of the study was to investigate the inhibition activities of polyisoprenoids in mangrove plant Avicennia marina in WiDr cells induces on P13k, Akt1, mammalian target of rapamycin (mTOR), Egfr, and P53 gene expression using reverse transcription-polymerase chain reaction (RT-PCR). MATERIAL AND METHODS: The leaves of A. marina were dried and extracted with n-hexane followed by evaporation and freeze-drying. Polyisoprenoid contents were analyzed with two-dimensional thin-layer chromatography method. Cell viability was assessed with 3-(4,5-dimetiltiazol-2-il)-2,5-diphenyl tetrazolium bromide assay. The cycle cell was tested with flow cytometry method. The apoptotic test was determined with a double-staining method. The gene expression on P13k, Akt1, mTOR, Egfr, and P53 was analyzed by RT-PCR method. RESULTS: The results showed that 48 h cytotoxic activity of polyisoprenoids against WiDr cells and 5-Fu (positive control) had IC50 values, 295.25 μg/mL and 17.43 μg/mL. Cell cycle analysis depicted that the inhibition of polyisoprenoid occurred in the G0-G1 phase and 5-Fu in S phase. Polyisoprenoid and 5-Fu had the same mechanism in the early apoptotic phase. RT-PCR revealed that polyisoprenoids downregulated the P13k, Akt1, mTOR, and Egfr gene expression, and however, upregulated P53 gene expression. CONCLUSION: The present study confirmed that polyisoprenoids from A. marina leaves showing as chemopreventive agents for colon cancer.


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