• BMB Reports
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• v.40 no.1
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• pp.22-28
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• 2007

MyoD, expressed in skeletal muscle lineages of vertebrate embryo, is one of muscle-specific basic helix-loop-helix (bHLH) transcription factors, which plays a key role in the determination and differentiation of all skeletal muscle lineages. In this study, a cDNA of grass carp MyoD was cloned and characterized from total RNA of grass carp embryos by RT-PCR. The full-length cDNA of grass carp MyoD is 1597 bp. The cDNA sequence analysis reveals an open reading frame of 825 bp coding for a protein of 275 amino acids, which includes a bHLH domain composed of basic domain (1-84th amino acids) and HLH domain (98-142th amino acids), without signal peptide. Then the MyoD cDNA of grass carp was cloned to yeast expression vector pPICZ$\alpha$A and transformed into P. pastoris GS115 strain, the recombinant MyoD protein with a molecular weight of about 31KD was obtained after inducing for 2d with 0.5% methanol in pH 8.0 BMGY medium, and the maximum yield was about 250 mg/L in shaking-flask fermentation. The results were expected to benefit for further studies on the crystal structure and physiological function of fish MyoD.

• Tuberculosis and Respiratory Diseases
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• v.86 no.4
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• pp.304-318
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• 2023

Background: Cancer-associated fibroblasts (CAFs) are key components of the tumor microenvironment and significantly contribute to immune evasion. We investigated the effects of CAFs on the immune function of CD4+ and CD8+ T cells in non-small cell lung cancer (NSCLC). Methods: We isolated CAFs and normal fibroblasts (NFs) from tumors and normal lung tissues of NSCLC patients, respectively. CAFs were co-cultured with activated T cells to evaluate their immune regulatory function. We investigated the effect of CAF conditioned medium (CAF-CM) on the cytotoxicity of T cells. CAFs were also co-cultured with activated peripheral blood mononuclear cells and further incubated with cyclooxygenase-2 (COX2) inhibitors to investigate the potential role of COX2 in immune evasion. Results: CAFs and NFs were isolated from the lung tissues (n=8) and lymph nodes (n=3) of NSCLC patients. Immune suppressive markers, such as COX2 and programmed death-ligand 1 (PD-L1), were increased in CAFs after co-culture with activated T cells. Interestingly, CAFs promoted the expression of programmed death-1 in CD4+ and CD8+ T cells, and strongly inhibited T cell proliferation in allogenic and autologous pairs of CAFs and T cells. CAF-CM decreased the cytotoxicity of T cells. COX2 inhibitors partially restored the proliferation of CD4+ and CD8+ T cells, and downregulated the expression of COX2, prostaglandin E synthase, prostaglandin E2, and PD-L1 in CAFs. Conclusion: CAFs promote immune evasion by suppressing the function of CD4+ and CD8+ T cells via their effects on COX2 and PD-L1 in NSCLC. The immunosuppressive function of CAFs could be alleviated by COX2 inhibitors.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.8
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• pp.3829-3833
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• 2016

Inhibition of cancer-associated fibroblasts (CAFs) may improve the efficacy of cancer therapy. Polysaccharide extracted from polygonatum can selectively inhibit the growth of prostate-CAFs (p<0.001) without inhibiting the growth of normal fibroblasts (NAFs). Polysaccharides from polygonatum stimulate autophagy of prostate-CAFs. 3-methyl-adenine(3-MA) is an autophagy inhibitor. 3-MA was added to prostate-CAFs with polysaccharide from polygonatum to determine whether autophagy plays an important role in the restrained effect. Finally, polysaccharide from polygonatum treatment significantly increased the activation of Beclin-1 and LC3, key autophagy proteins. Polysaccharide from polygonatum stimulates autophagy of prostate-CAFs and inhibits prostate-CAF growth, indicating that a novel anti-cancer strategy involves inhibiting the growth of prostate-CAFs.

• Journal of dental hygiene science
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• v.21 no.4
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• pp.227-232
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• 2021

Background: Cancer-associated fibroblasts (CAFs) are abundant in tumor microenvironments and interact with cancer cells to promote tumor proliferation in oral squamous cell carcinoma (OSCC). Cathepsin D (CTSD) is a soluble lysosomal aspartic endopeptidase involved in tumor proliferation and angiogenesis. In this preliminary study, we observed CTSD expression in OSCC and CAFs, postulating that CTSD might act as a bridge between OSCC and CAFs. Methods: Human epidermal keratinocytes (HEKs), OSCC, and immortalized human normal oral fibroblasts (hTERT-hNOFs) were used in this study. Additionally, we used hTERT-hNOFs transfected with an empty vector, WT (wild-type)-YAP (Yes-associated protein), and YAPS127A (YAP serine 127 to alanine). YAP127A hTERT-hNOFs activated fibroblasts similar to CAFs. To identify CTSD expression between OSCC and CAFs, conditioned medium (CM) was collected from each cell. Protein expression of CTSD was identified by western blotting. Results: To identify the expression of CTSD in fibroblasts stimulated by OSCC, we treated fibroblasts with CM from HEK and OSCC. Results indicated that hTERT-hNOFs with OSCC CM showed a weakly increased expression of CTSD compared to stimulation by HEK CM. This indicates that CAFs, YAPS127 hTRET-hNOFs, overexpress CTSD protein. HEK cells showed no CTSD expression, regardless of treatment with fibroblast CM, whereas OSCC highly expressed CTSD proteins compared with the CTSD expression in HEK cells. We also found that CTSD expression was unaffected by changes in transforming growth factor-β levels. Conclusion: This study proposes that CTSD might have potential as an interacting executor between OSCC and CAFs. Further studies are needed to investigate the role of CTSD in tumor and stromal cells.

• Proceedings of the Korean Society of Crop Science Conference
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• 2022.10a
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• pp.32-32
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• 2022

Abaca (Musa textilis L. Nee) is a native Musa species from the Philippines known for its natural fiber. Abaca fiber a.k.a. Manila hemp extracted from its pseudostems is considered one of the strongest fibers in the world. This is used for commodities such as ropes, papers, and money bills. Abaca is vulnerable to pests and diseases such as the Abaca Bunchy Top Disease (ABTD) caused by Abaca Bunchy Top Virus (ABTV) and Banana Bunchy Top Virus (BBTV). Inosa, one of the varieties of abaca utilized in the Philippines, is highly susceptible to ABTD. In contrast, Pacol (Musa balbisiana L.), a close relative of abaca, is highly resistant to the same disease. Here, we report the sequencing and de novo genome assembly of both abaca var. Inosa and banana var. Pacol. A total of ~16 Gb and ~21 Gb raw reads for Inosa and Pacol, respectively, were generated using Pacbio Hifi sequencing method and assembled with Hifiasm. High-quality de novo assemblies of both Musa species with 99% recovered as per BUSCO analysis were obtained. The assembled Inosa genome has a total length of ~654 Mb and N50 of 7 Mb while Pacol has a total length of 527 Mb and N50 of 3 Mb which are close to their estimated genome size of ~638 Mb and ~503 Mb, respectively. The information that can be derived from the de novo assembled genomes would provide a solid foundation for further research in disease resistance and fiber quality improvement in abaca.

• Journal of Periodontal and Implant Science
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• v.46 no.1
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• pp.22-34
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• 2016

Purpose: Gingival recession is a major esthetic concern and may lead to root sensitivity during periodontal treatment. Coronally advanced flaps (CAFs) with and without acellular dermal matrix (ADM) are widely used in root coverage procedures. The aim of this study was to analyze the efficacy of CAF in combination with ADM in the treatment of gingival recession. Methods: PubMed, The Cochrane Library, and Embase were used to identify relevant articles. The articles were screened, data were extracted, and the quality of the studies was assessed by three reviewers with expertise in clinical practice, trials, statistics, and biomedical editing. The clinical endpoints of interest included changes in recession, probing depth (PD), clinical attachment level (CAL), and keratinized tissue (KT). Results: Ten randomized controlled trials were identified, including six studies that compared CAFs with ADM and CAFs using connective tissue grafting (CTG) and four studies that compared CAFs with or without ADM. No statistically significant differences were found between the use of ADM and CTG, whereas statistically significant differences were found between groups in which ADM and CAF were combined and groups that underwent CAF alone with regard to recession coverage, CAL, and KT. The combination of CAF with an ADM allograft achieved more favorable recession coverage and recovery of CAL and KT than CAF alone. Conclusions: The results from the ADM and CTG groups suggest that both procedures may be equally effective in clinical practice. Given the limitations of this study, further investigation is needed to clarify the effectiveness of ADM and CAF in clinical practice.

• Biomolecules & Therapeutics
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• v.32 no.3
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• pp.281-290
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• 2024

Pancreatic ductal adenocarcinoma (PDAC) has a poor prognosis owing to its desmoplastic stroma. Therefore, therapeutic strategies targeting this tumor stroma should be developed. In this study, we describe the heterogeneity of cancer-associated fibroblasts (CAFs) and their diverse roles in the progression, immune evasion, and resistance to treatment of PDAC. We subclassified the spatial distribution and functional activity of CAFs to highlight their effects on prognosis and drug delivery. Extracellular matrix components such as collagen and hyaluronan are described for their roles in tumor behavior and treatment outcomes, implying their potential as therapeutic targets. We also discussed the roles of extracellular matrix (ECM) including matrix metalloproteinases and tissue inhibitors in PDAC progression. Finally, we explored the role of the adaptive and innate immune systems in shaping the PDAC microenvironment and potential therapeutic strategies, with a focus on immune cell subsets, cytokines, and immunosuppressive mechanisms. These insights provide a comprehensive understanding of PDAC and pave the way for the development of prognostic markers and therapeutic interventions.

• Oncology Letters
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• v.18 no.5
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• pp.4645-4650
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• 2019

Tumor microenvironment serves an important role in tumor growth and metastasis. Cancer cells can promote growth and malignancy by altering the surrounding stroma. Cancer-associated fibroblast (CAF) are an abundant cell type present within the tumor microenvironment and provide tumorigenic features by secreting cytokines. In the current study, the CAF-mediated invasion of oral squamous cell carcinoma (OSCC) was investigated and the associated mechanisms were elucidated. Cancer invasion was estimated using a Matrigel-coated Transwell chamber and FITC-gelatin matrix. To verify the effect of the tumor microenvironment, conditioned media (CM) from normal fibroblast (NF) and CAFs were prepared. An ELISA was performed to estimate the level of IL-1β. A proteome profiler human protease array was performed to verify the proteases affected by stimulation with CM, from CAF. Recombinant IL-1β protein increased the invasion of OSCC cells. IL-1β expression was higher in CAF than NF. CM from CAF (CM-CAF) increased cancer invasion and FITC-gelatin matrix degradation. The invasive capacity provided by CAF was abrogated by an IL-1 receptor (IL-1R) antagonist. Additionally, CM-CAF increased the secretion of ADAM 9 and Kallikrein 11 from OSCC cells. The invasion activity by CM-CAF was partially abrogated by the neutralization of ADAM 9 or Kallikrein 11. In conclusion, by providing stromal factor, CAFs were a critical inducer of OSCC invasion, and CAF secretes the required amount of IL-1β to increase cancer invasion activity. The invasive capacity of CAF was identified to be IL-1R-dependent. ADAM 9 and Kallikrein 11 were influencing factors involved in the increase of CAF-mediated cancer invasion.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.7
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• pp.3043-3050
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• 2015

The tumor microenvironment (TME) includes numerous non-neoplastic cells such as leukocytes and fibroblasts that surround the neoplasm and influence its growth. Tumor-associated macrophages (TAMs) and cancerassociated fibroblasts (CAFs) are documented as key players in facilitating cancer appearance and progression. Alteration of the macrophage (CD68, CD163) and fibroblast (${\alpha}-SMA$, FSP-1) cells in Opisthorchis viverrini (Ov) -induced cholangiocarcinoma (CCA) was here assessed using liver tissues from an established hamster model and from 43 human cases using immunohistochemistry. We further investigated whether M2-activated TAMs influence CCA cell migration ability by wound healing assay and Western blot analysis. Macrophages and fibroblasts change their phenotypes to M2-TAMs (CD68+, CD163+) and CAFs (${\alpha}-SMA+$, FSP-1+), respectively in the early stages of carcinogenesis. Interestingly, a high density of the M2-TAMs CCA in patients is significantly associated with the presence of extrahepatic metastases (p=0.021). Similarly, CD163+ CCA cells are correlated with metastases (p=0.002), and they may be representative of an epithelial-to-mesenchymal transition (EMT) with increased metastatic activity. We further showed that M2-TAM conditioned medium can induce CCA cell migration as well as increase N-cadherin expression (mesenchymal marker). The present work revealed that significant TME changes occur at an early stage of Ov-induced carcinogenesis and that M2-TAMs are key factors contributing to CCA metastasis, possibly via EMT processes.

• Korean Journal of Head & Neck Oncology
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• v.30 no.2
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• pp.43-52
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• 2014

배경 및 목적 암-연관 섬유아세포(Cancer-associated fibroblasts, CAF)는 종양미세환경의 가장 중요한 요소의 하나다. 그래서 두경부편평세포암에 대해 CAF가 암세포의 운동성에 미치는 영향을 평가하고, CAF에 과발현되는 CD44의 역할에 대해 평가하고자 하였다. 재료 및 방법 두경부암환자의 종양조직에서 CAF를 분리하고, 비종양성 조직으로부터 정상 섬유아세포(NHF)를 분리하였다. 창상치유분석과 상하 챔버를 이용한 3차원 세포 이동 분석을 이용하여, CAF가 암세포의 이동에 미치는 영향을 분석하고, CAF에서 과발현되는 CD44를 중화항체로 CAF를 차단했을 때 암세포 이동의 변화를 관찰하였다. 결과 NHF에 비해 CAF에서 CD44가 과발현되는 것을 관찰하였다. 창상치유분석에서 CAF와 같이 배양된 암세포는 NHF와 같이 배양된 암세포에 비해 더 빠른 이동을 보였다. CD44 중화 항체를 처리했을 때는 암세포의 이동성이 저해되었다. 결론 CAF는 종양미세환경에서 암세포의 운동성을 조절하는 중요한 인자의 하나일 것으로 사료된다. CD44는 CAF의 기능을 매개하는 중요한 표지자 중 하나로 생각된다.