scholarly journals Molecular Pharming of the Recombinant Protein hEGF-hEGF Concatenated with Oleosin Using Transgenic Arabidopsis

Genes ◽  
2020 ◽  
Vol 11 (9) ◽  
pp. 959
Author(s):  
Weidong Qiang ◽  
Tingting Gao ◽  
Xinxin Lan ◽  
Jinnan Guo ◽  
Muhammad Noman ◽  
...  

We set out to assess the NIH/3T3 cell proliferation activity of Arabidopsis oil body-expressed recombinant oleosin–hEGF–hEGF protein. Normally, human epidermal growth factor (hEGF) is purified through complex process, however, oleosin fusion technology provides an inexpensive and scalable platform for its purification. Under a phaseolin promoter, we concatenated oleosin gene to double hEGF (hEGF–hEGF) with plant-preferred codons in the expression vectors and the construct was transformed into Arabidopsis thaliana (Arabidopsis). The transgenic Arabidopsis was validated by RT–PCR and the content of recombinant protein oleosin–hEGF–hEGF was quantified by western blot. Subsequently, the proliferation assay and transdermal absorption were determined by MTT method and immunohistochemical staining, respectively. First, the expression level of hEGF was recorded to be 14.83-ng/μL oil body and due to smaller size transgenic oil bodies expressing the recombinant oleosin–hEGF–hEGF, they were more skin permeable than those of control. Second, via the staining intensity of transgenic oil bodies was greater than EGF at all time points via immunohistochemical staining in transdermal absorption process. Lastly, activity assays of oil bodies expressed oleosin–hEGF–hEGF indicated that they stimulated the NIH/3T3 cell proliferation activity. Our results revealed oil-body-expressed oleosin–hEGF–hEGF was potential new material having implications in the field of medicine.

1970 ◽  
Vol 7 (3) ◽  
pp. 14-19 ◽  
Author(s):  
Hekdin Marsius Sipayung ◽  
Jansen Silalahi ◽  
Yuandani Y

Objectives: The objective of this study was to investigate the activity of combination of hydrolyzed VCO (HVCO) and chitosan on NIH 3T3 cell proliferation activity, NIH 3T3 cell migration, COX-2 and VEGF protein expression. Design: In vitro cytotoxic assay was determined by MTT (MicrocultureTetrazoliumTehnique) assay, cell proliferation activity was measured by calculating cell viability incubated 24 hours, 48 hours and 72 hours, wound closure percentage was tested by scratch wound healing method, expression of COX-2 protein and VEGF protein were measured by immunocytochemical method. Interventions: The variable that was intervened in this study was the concentration of HVCO and chitosan. Main Outcome Measures: The main measurements carried out in this study were the absorbance value of HVCO and chitosan which was converted into viability cell, proliferation activity, percentage of wound closure, and percentage of COX-2 and VEGF protein expression. Results: Cytotoxic activity of HVCO and chitosan resulted the best concentration at 31.25 μg/ml, scratch wound healing assay from a combination HVCO and chitosan resulted the best migration of fibroblast cells at a ratio of 1:1 with HVCO 62.5 μg/ml and chitosan 62.5 μg/ml, combination of HVCO 62.5 μg/ml and chitosan 62.5 μg/ml (1:1) increased expression of COX-2 and VEGF. Conclusion: Combination of HVCO and chitosan could increase NIH 3T3 cell migration, COX-2 and VEGF protein expression. Combination of HVCO and chitosan had better wound healing activity in vitro than single use. Keywords: Rhizomucor miehei, viability, proliferation, migration, expression


1999 ◽  
Vol 188 (3) ◽  
pp. 289-293 ◽  
Author(s):  
Satu-Leena Sallinen ◽  
Pauli K. Sallinen ◽  
Juha T. Kononen ◽  
Kirsi M. Syrj�koski ◽  
Nina N. Nupponen ◽  
...  

Author(s):  
Suriya Kumaresan ◽  
Rema Ramasamy ◽  
Philip Robinson Jayachandran

Objectives: Medicinal plants and herbs are used in combination in Ayurveda and folklore medicine as they exhibit good cytotoxic activity. In the present study, the antioxidant, phytochemical, and cell proliferation activity of the combined crude methanolic extract of Ficus religiosa and Ficus benghalensis leaves were investigated.Methods: Antioxidant activity was performed by 2, 2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) and hydrogen peroxide methods, and the presence of the phytochemicals was screened using the gas chromatography–mass spectrometry. The extract was further evaluated for its cell proliferation activity against cancer cells using the mitochondrial reduction assay. Antioxidant property of the extracts was measured using the DPPH, hydrogen peroxide, and ferric-reducing antioxidant power assay, respectively, using the UV spectrophotometer.Results: The combined extract exhibited strong antioxidant potential in DPPH assay by increase in the percentage of inhibition with the increase in concentration. Similarly, the IC50 value of the methanol extract in peroxidase scavenging activity was 49.85 μg/mL comparatively lower than the ascorbic acid used as standard. The phytochemical analysis of the methanol extract showed the presence of nine phytoconstituents, which exhibit antioxidant and anticancer property. Mitochondrial reduction assay performed to evaluate the cell proliferation activity of the combined leave extract showed that increase in the concentration of the extract decreased the cell proliferation in the HeLa cell line.Conclusion: The results of present study show a possible synergistic activity of leaves against human cervical cancer.


2007 ◽  
Vol 17 (3) ◽  
pp. 607-614 ◽  
Author(s):  
A. YOSHIDA ◽  
L.O. SARIAN ◽  
L.A.L.A. ANDRADE ◽  
F. PIGNATARO ◽  
G.A. PINTO ◽  
...  

Sign in / Sign up

Export Citation Format

Share Document