Cell and Tissue Journal

Abstract Aim: In this study, the effects of recombinant human growth hormone and Gemcitabine alone and in combination with each other were investigated on the lung fibroblast cell lines.
Material and methods: Human growth hormone at 10-400 ng/ml and Gemcitabine at 1-100 μg/ml concentrations were treated with the human lung fibroblast cells and the analysis were performed with MTT assay, propidium iodide staining and scratch assay.
Results: Studies of the cell cycle and MTT assay revealed the effect of growth hormone on the progression of the cell cycle and exit of G1 phase and the cell proliferation, and the inhibitory effects of the cell cycle and growth process by Gemcitabin compared with the control sample. The results of the scratch test indicated that the growth hormone at its effective concentrations caused increased the cell migration compared to control, while the Gemcitabin reduced the migration of the cell to the control.
Conclusions: Using growth hormone in combination to gemcitabin during chemotherapy treatment, it seems that we can reduce the rate of the neem normal cells death. Since these cells are more sensitive to growth hormone´s effect than cancer cells.
 

Abstract Aim: In this century, the production of recombinant drugs such as growth hormone has increased. Different problems existed in the expressions of cytoplasmic and Periplasmic types of growth hormone. Therefore, finding a way to optimize expression is very necessary. In this study, we optimized expression of growth hormone in the form of solution state by trx-tag method. This method increase protein expression (Periplasmic problem) and also prevents formation of inclusion body (problem cytoplasmic).
Material and methods: Gene synthesis and gene cassette was done in pET 32a expression vector. Gene cassette contains trx tag for protein solubilization, His tag for purification and enterokinase for separate rHgh from previous tags.
 Results: After cloning the gene in vector, its expression was confirmed by Western blot technique. The results showed that the expression of fusion protein was done well.
Conclusion: the obtained finding proved that protein can be soluble by Trx-tag and increased its expression levels. Moreover, better results can be achieved in the fermentation and downstream processing.