• Korean Chemical Engineering Research
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• v.56 no.5
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• pp.711-717
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• 2018

Human epidermal growth factor (hEGF) can stimulate the division of various cell types and has potential clinical applications. Since the protein contains three intra-molecular disulfide bonds, the high expression of active hEGF in Escherichia coli has not been well researched, We fused the hEGF gene with a small ubiquitin-related modifier gene (SUMO) by synthesizing an artificial SUMO-hEGF fusion gene that was highly expressed in E. coli (DE3) strain. The optimal expression level of the soluble fusion protein, SUMO-hEGF with IPTG (Isopropyl-${\beta}$-D-Thiogalactopyranoside), was up to 38.9% of the total cellular protein. The fusion protein was purified by Ni-NTA affinity chromatography and cleaved by a SUMO-specific protease to obtain the native hEGF, which was further purified by Ni-NTA affinity chromatography. The result of the reverse-phase HPLC showed that the purity of the recombinant cleaved hEGF was greater than 98%.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.2
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• pp.149-154
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• 2005

Secretion of the expressed heterologous proteins can reduce the stress to the host cells and is beneficial to their recovery and purification. In this study, fed-batch cultures of Escherichia coli YK537 (pAET-8) were conducted in a 5-L fermentor for the secretory production of human epidermal growth factor (hEGF) whose expression was under the control of alkaline phosphatase promoter. The effects of feeding of glucose and complex nitrogen sources on hEGF production were investigated. When the fed-batch culture was conducted in a chemically de-fined medium, the cell density was 9.68 g/L and the secreted hEGF was 44.7 mg/L in a period of 60 h. When a complex medium was used and glucose was added in pH-stat mode, the secreted hEGF was improved to 345 mg/L. When the culture was fed with glucose at a constant specific rate of $0.25\;gg^{-1}h^{-1}$, hEGF reached 514 mg/L. The effects of adding a solution containing yeast extract and tryptone were further studied. Different rate of the nitrogen source feeding resulted in different levels of phosphate and acetic acid formation, thus affected hEGF expression. At the optimal feeding rate, hEGF production achieved 686 mg/L.

• Journal of Ginseng Research
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• v.43 no.4
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• pp.550-561
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• 2019

Background: Gastric ulcer (GU) is a common gastrointestinal disease that can be induced by many factors. Finding an effective treatment method that contains fewer side effects is important. 20 (S)-ginsenoside Rg3 is a kind of protopanaxadiol and has shown superior antiinflammatory and antioxidant effects in many studies, especially cancer studies. In this study, we examined the treatment efficacy of 20 (S)-ginsenoside Rg3 on GU. Methods: Three kinds of GU models, including an alcohol GU model, a pylorus-ligated GU model, and an acetic acid GU model, were used. Mouse endothelin-1 (ET-1) and nitric oxide (NO) levels in blood and epidermal growth factor (EGF), superoxide dismutase, and NO levels in gastric mucosa were evaluated. Hematoxylin and eosin staining of gastric mucosa and immunohistochemical staining of ET-1, inducible nitric oxide synthase (NOS2), and epidermal growth factor receptors were studied. Ulcer index (UI) scores and UI ratios were also analyzed to demonstrate the GU conditions in different groups. Furthermore, Glide XP from $Schr{\ddot{o}}dinger$ was used for molecular docking to clarify the interactions between 20 (S)-ginsenoside Rg3 and EGF and NOS2. Results: 20 (S)-ginsenoside Rg3 significantly decreased the UI scores and UI ratios in all the three GU models, and it demonstrated antiulcer effects by decreasing the ET-1 and NOS2 levels and increasing the NO, superoxide dismutase, EGF, and epidermal growth factor receptor levels. In addition, high-dose 20 (S)-ginsenoside Rg3 showed satisfactory gastric mucosa protection effects. Conclusion: 20 (S)-ginsenoside Rg3 can inhibit the formation of GU and may be a potential therapeutic agent for GU.

• Korean Journal of Agricultural Science
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• v.47 no.4
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• pp.815-827
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• 2020

The epidermal growth factor (EGF) transgenic soybean was developed and biosynthesis of human epidermal growth factor (hEGF) in soybean seeds was confirmed. Also, EGF transgenic soybean were found to contain a herbicide resistance selectable marker by introduction of phosphinothricin acetyltransferase (PAT) gene from the Streptomyces hygroscopicus. For biosafety assessment, the EGF transgenic soybean expressing the EGF biosynthesis gene EGF and herbicide resistant gene PAT was tested to determine effects on survival of Cyprinus carpio, commonly used as a model organism in ecotoxicological studies. C. carpio was fed 100% ground soybean suspension, EGF soybean or non-genetically modified (GM) counterpart soybean (Gwangan). Gene expression of EGF soybean was confirmed by PCR and ELISA to have EGF/PAT. Feeding test showed that no significant differences in cumulative immobility or abnormal response between C. carpio samples fed on EGF soybean and non-GM counterpart soybean. The 48 h-EC50 values of the EGF and non-GM soybean were 1,688 mg·L-1 (95% confidence limits: 1,585 - 1,798 mg·L-1) and 1,575 mg·L-1 (95% confidence limits: 1,433 - 1,731 mg·L-1), respectively. The soybean NOEC (no observed effect concentration) value for C. carpio was suggested to be 625 mg·L-1. We concluded that there was no significant difference in toxicity for non-target organisms (C. carpio) between the EGF soybean and non-GM counterparts.

• Korean Journal of Agricultural Science
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• v.48 no.4
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• pp.1003-1013
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• 2021

Thioredoxin (TRX) protein is an antioxidant responsible for reducing other proteins by exchanging cysteine thiol-disulfide and is also known for its anti-allergic and anti-aging properties. On the other hand, epidermal growth factor (EGF) is an important material used in the cosmetics industry and an essential protein necessary for dermal wound healing facilitated by the proliferation and migration of keratinocytes. EGF also assists in the formation of granulation tissues and stimulates the motility of fibroblasts. Hence, genetically modified soybeans were developed to overexpress these industrially important proteins for mass production. A single-dose oral-toxicity-based study was conducted to evaluate the potential toxic effects of TRX and EGF proteins, as safety assessments are necessary for the commercial use of seed-specific protein-expressing transgenic soybeans. To achieve this rationale, TRX and EGF proteins were mass purified from recombinant E. coli. The single-dose oral-toxicity tests of the TRX and EGF proteins were carried out in six-week old male and female Institute of Cancer Research (ICR) mice. The initial evaluation of the single-dose TRF and EGF treatments was based on monitoring the toxicity signatures and mortality rates among the mice, and the resultant mortality rates did not show any specific clinical symptoms related to the proteins. Furthermore, no significant differences were observed in the weights between the treatment and control groups of male and female ICR mice. After 14 days of treatment, no differences were observed in the autopsy reports between the various treatment and control groups. These results suggest that the minimum lethal dose of TRX and EGF proteins is higher than the allowed 2,000 mg·kg^-1 limit.

• Journal of Korean Dental Science
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• v.16 no.2
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• pp.172-181
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• 2023

Purpose: To investigate the effect of epidermal growth factor (EGF) with collagen matrix (CM) for increasing gingival thickness. Materials and Methods: In five mongrel dogs, bilateral gingival defects were surgically made on the maxillary canines. After two months, either a subepithelial connective tissue graft (group SCTG) or CM with EGF (0.1 ug/ml, group EGF) was grafted, and the flap was coronally positioned to cover the graft materials. The animals were sacrificed after three months. Intraoral scanning was performed for soft tissue analysis. Histologic and histomorphometric analyses were performed. Result: Two animals exhibited wound dehiscence during the healing phase, leaving three for analysis. No statistically significant difference was found in soft tissue changes (P>0.05). The level of gingival margin (GM) increased in both groups (1.02±0.74 mm in group SCTG vs. 1.24±0.83 mm in group EGF). Linear increases at the GM pre-augmentation in the soft tissue profile were 1.08±0.58 mm in group SCTG and 0.96±0.73 mm in group EGF. Histomorphometric parameters (keratinized tissue height, tissue thickness, and rete peg density) were not significantly different between the groups (P>0.05). Conclusion: EGF loaded onto CM led to comparable gingival phenotype enhancement to SCTG.

• Korean Journal of Radiology
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• v.23 no.9
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• pp.921-930
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• 2022

Objective: To identify epidermal growth factor receptor (EGFR) mutations in lung adenocarcinoma based on ¹⁸F-fluorodeoxyglucose (FDG) PET/CT radiomics and clinical features and to distinguish EGFR exon 19 deletion (19 del) and exon 21 L858R missense (21 L858R) mutations using FDG PET/CT radiomics. Materials and Methods: We retrospectively analyzed 179 patients with lung adenocarcinoma. They were randomly assigned to training (n = 125) and testing (n = 54) cohorts in a 7:3 ratio. A total of 2632 radiomics features were extracted from the tumor region of interest from the PET (1316) and CT (1316) images. Six PET/CT radiomics features that remained after the feature selection step were used to calculate the radiomics model score (rad-score). Subsequently, a combined clinical and radiomics model was constructed based on sex, smoking history, tumor diameter, and rad-score. The performance of the combined model in identifying EGFR mutations was assessed using a receiver operating characteristic (ROC) curve. Furthermore, in a subsample of 99 patients, a PET/CT radiomics model for distinguishing 19 del and 21 L858R EGFR mutational subtypes was established, and its performance was evaluated. Results: The area under the ROC curve (AUROC) and accuracy of the combined clinical and PET/CT radiomics models were 0.882 and 81.6%, respectively, in the training cohort and 0.837 and 74.1%, respectively, in the testing cohort. The AUROC and accuracy of the radiomics model for distinguishing between 19 del and 21 L858R EGFR mutational subtypes were 0.708 and 66.7%, respectively, in the training cohort and 0.652 and 56.7%, respectively, in the testing cohort. Conclusion: The combined clinical and PET/CT radiomics model could identify the EGFR mutational status in lung adenocarcinoma with moderate accuracy. However, distinguishing between EGFR 19 del and 21 L858R mutational subtypes was more challenging using PET/CT radiomics.

• Reproductive and Developmental Biology
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• v.30 no.3
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• pp.163-167
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• 2006

The purpose of the present study was to establish culture conditions for the in vitro study of the neonatal piglet Sertoli cell. Isolation for the culture of Sertoli cell was established using collagenase and pancreatin digestion of testicular tissues. The effects of various culture media, fetal bovine serum(FBS), follicular stimulating hormone(FSH), epidermal growth factor(EGF) and insulin-transferrin-sodium selenite(ITS) on growth of neonatal piglet Sertoli cells were investigated. The mitogenic effects of Dulbecco's modified Eagle's medium+Ham's F-12 medium was higher than other media used in this experiment. The addition of 1% FBS in cultures was necessary for attachment of Sertoli cell clusters. However, except FBS and EGF, FSH and ITS did not stimulate Sertoli cell proliferation. When Sertoli cells isolated from neonatal piglets were cultured in Dulbecco's modified Eagle's medium+Ham's F-12 medium supplemented with 1% FBS, FSH EGF and ITS, the yield and plating efficiency of Sertoli cells were largely increased. Confluency of Sertoli cells was reached as early as 4 days of culture. The method described here reduces or eliminates many of the drawbacks of the conventional procedures used to isolate and culture of Sertoli cells, thus providing a useful tool in studies of growth kinetics and regulation of cell proliferation in vitro.

• Clinical and Experimental Reproductive Medicine
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• v.23 no.3
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• pp.303-310
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• 1996

The objective of this study was to examine the effect of embryos development following IVF of in vitro-matured porcine oocytes treated with epidermal growth factor (EGF). When cumulus-enclosed oocytes were incubated in TCM 199 medium supplemented with (1) control group, (2) 10 ng/ml EGF, (3) 10${\mu}g$ml FSH and 10% FBS, or (4) 10 ng/ml EGF, 10 ${\mu}g$/ml FSH, and 10% FBS for 42 hr, the late developmental rates on NCSU (0.4% BSA) medium after fertilization were higher in (3) and (4) groups (13.4, 18.3%) than in (2) group (5.2%, p < 0.005), but (2) group is significantly higher than the development to blastocyst of oocytes of (1) group (1.2%). Also, when the cell number of total, ICM, and TE of those blastocysts at 6 day produced in vitro was investigated by double staining (PI and bisbenzimide), total cell number of (4) group (58.80${\pm}$ 11.90) was higher than that of (2) and (3) groups (42.17${\pm}$9.97, 49.07${\pm}$9.77, P < 0.05). ICM cell number of blastocysts of (4) group (11.69${\pm}$5.56) was higher than that of (2) and (3) groups (5.00${\pm}$4.24, 6.77${\pm}$4. 92, P < 0.05). Furthermore, the proportion of ICM in (4) group (19.0${\pm}$1.6) was higher than that in (2) and (3) groups (11.1${\pm}$3.0, 12. 7${\pm}$2.1). These results suggested that in vitromatured porcine oocytes treated with EGF alone can be developed to blastocyst, but high proportion on the development to blastocyst and number of total cell and ICM in blastocyst can be obtained when supplemented with additional FSH and FBS.

• Korean Journal of Veterinary Research
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• v.43 no.2
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• pp.211-218
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• 2003

Human ovarian cancerous cells survive in a way that they trigger the nucleotide excision repair (NER) or double-strand DNA repair (dsDNA) repair mechanism to show resistance to anticancer drugs and activate many kinds of repair protein, thus removing damaged DNAs. Two experiments on the PA-1 human ovarian teratocareinoma cell line that hardly has any expression of epidermal growth factor receptor (EGFR) were conducted in the study; first, EGF-R was transfected and its receptor was obtained. The receptor was investigated in terms of its mutual relations with many kinds of protein concerning NER or dsDNA repair. Second, it was examined what kind impact cisplatin and adriamycin had on the effects of EGF-R over the PA-1 cell line lacking EGF-R. When being administered with cisplatin and adriamycin, Hey and Hey C2 cell lines showed a high level of resistance while PA-1 cell line a high level of sensitivity. Hey and Hey C2 cell lines that are resistant against anticancer drugs exhibited a high level of EGF-R expression while PA-1 cell line that is sensitive to them did a much lower level of the expression. When PA-1 cell line was transfected for the expression of DNA adduct and EGF-R, it showed a higher level of resistance compared to the control group. There was no difference in the expression of DNA repair proteins (DNA- dependent protein kinase, Ku70, and Ku80) between Hey and the PA-1 cell lines. The results indicate that the Hey cell line that is resistant against cisplatin and adriamycin works along the signaling system responding to the changes of EGF-R while the PA-1 cell line that is sensitive to both of them does to the lack of EGF-R.