Salivary gland adenocarcinoma(AdCa NOS) is one of the major causes of mortality among malignant salivary gland tumors. New therapeutic measure are needed to improve the outcome for patients with AdCa NOS because current therapy does not significantly improve survival rates. Transglutaminase 2(TGase 2) was implicated in forming cross-linked protein polymer, apoptosis and matrix interaction. And also TGase 2 expression is up-regulated in proliferation, migration, invasion, and metastasis of cancer cells. shRNA which has emerged as an effective method to target specific genes for silencing has provided new opportunities for cancer therapy. But there has been rarely reported using shRNA-TGase 2 transfection in AdCa NOS. The purpose of this study were to examine the specific inhibition of TGase 2 mRNA and protein expression by siRNA transfection of TGase 2 through RT-PCR and immunoslot blotting, and to study proliferation, migration and invasion assay of SGT cell line from AdCa NOS. Cell cycle analysis showed that the downregulation of shRNA-TGase 2 caused the accumulation of cells in the sub-G0/G1 phase. In migration assay, suppressing shRNA-TGase 2 inhibited the capacity of the cells to migrate compared to parental cells. In invasion assay, cells transfected with shRNA-TGase 2 decreased in invasion when compared to SGT and vector transfected cells. shRNA-TGase 2 expressing plasmids efficiently downregulated TGase 2 mRNA and TGase 2 protein expression. It suggested that the shRNA-TGase 2 targeting system against TGase 2 could have a therapeutic potentiality for malignant salivary gland tumors, especially in inhibiting and/or preventing cancer cell proliferation, migration and invasion.
Vinpocetine induces anti-inflammatory effects in various inflammatory diseases via the inhibition of phosphodiesterase type-1-independent nuclear factor-κB signaling pathway and the release of inflammatory cytokines. In this study, we investigated the effect of vinpocetine on the proliferation of colon cancer cells and its underlying molecular mechanisms. Our data showed that vinpocetine inhibits the viability and proliferation of colon cancer cells. Vinpocetine treatment induced cell death in HCT116 cells, which the percentages of sub-G1 phase were significantly increased, and the apoptosis-related genes were regulated after HCT116 cells were treated with vinpocetine. In sum, our findings indicated that vinpocetine could be a therapeutically useful candidate in the treatment of colon cancer.
This study aimed to examine the effect of a mild elevation in serum cholesterol level in a porcine coronary overstretch restenosis model using a balloon angioplasty catheter or drug-eluting coronary stent. Pigs were divided into two groups and were fed a commercial normal diet (CND, n = 4) or a high-fat diet (HFD, n = 4) for 5 weeks. Coronary overstretch injury by balloon angioplasty or stent implantation was induced in the left anterior descending and left circumflex artery after 1 week of feeding. Histopathological analysis was performed at 4 weeks after coronary injury. During the experiment, the total cholesterol level in the HFD group increased by approximately 44.9% (from 65.9 ± 3.21 mg/dL at baseline to 95.5 ± 9.94 mg/dL at 5 weeks). The lumen area in the CND group was reduced in comparison with that in the HFD group after balloon angioplasty. After stent implantation, the injury score showed no significant difference. There were significant differences in the neointimal area (2.7 ± 0.33 mm2 in the CND group vs. 3.3 ± 0.34 mm2 in the HFD group, p<0.05), lumen area (2.6 ± 0.54 mm2 in the CND group vs. 2.0 ± 0.33 mm2 in the HFD group, p<0.05), and percent area stenosis (52.0 ± 7.96% in the CND group vs. 62.4 ± 5.15% in the HFD group, p<0.05). Body weight change was not different between the two groups. Increased serum cholesterol level activated vascular smooth muscle cell proliferation in the porcine coronary overstretch model.
The purpose of this study was to investigate the inhibitory effect of enzyme activity and anti-proliferation of human cancer cell lines (HCT 116, NCI-H460 and MCF-7) of peanut skin depending on cultivars (Arachis hypogaea L. cv. K-Ol, cv. Sinpalkwang, cv. Daan, cv. Heuksaeng) and extraction solvent. Peanut skin was extracted with 80% ethanol, 80% methanol, 80% acetone, and distilled water, followed by analysis of the enzyme inhibitory activity and anticancer activity. Methanol extract of Daan cultivar most effectively inhibited α-gluosidase (65.08%, 0.025 mg/mL), tyrosinase (82.49%, 2 mg/mL) and ACE (73.61%, 10 mg/mL). The inhibitory effect of peanut skin extracts on colon cancer cell (HCT-116), lung cancer cell (NCI-H460) and breast cancer cell (MCF-7) growth were investigate using MTT assay. The highest anti-proliferation of cancer cell line of peanut skin extracts was observed in the methanol extract of Daan cultivar. The cell viability on HCT 116, NCI-H460 and MCF-7 cell lines of methanol extracts from peanut skin of Daan cultivar was 48.13%, 41.03%, and 36.02% at 200 μg/mL, respectively. These results suggest that peanut skin extracts may mediate physiological activity, and provide valuable information for the use of peanut byproduct as a functional food material.
L-carnitine은 라이신과 메티오닌으로 생합성되며 골격근 과 심근을 포함한 다양한 동물조직에서 발견된다. L-carnitine이 포함된 식품으로는 양고기, 소고기, 돼지고기 등이 있고 근육발달에 도움을 주며 뼈를 강화하거나 대사작용을 도와주는 기능을 하여 영양 보조제로 많이 섭취하는 것으로 알려져 있다. 최근 L-carnitine은 제 2형 당뇨병, 골다 공증, 대사성 신경증후군 등의 다양한 질병의 약물로도 연구 되고 있으며 암에서는 치료 보조제로 개발되어있다. 하지만 대장암에서의 L-carnitine에 대한 효과 및 기전에 대해서는 명확하지 않고 연구된 바가 없기 때문에 본 연구에서 저자들은 L-carnitine의 효능을 인간대장암세포주 HCT116에서 규명하고자 하였다. L-carnitine은 세포 내 활성산소종 (ROS)를 높은 수준으로 증가시켜 세포 증식을 억제하였다. 또한, 세포 증식과 죽음에 관련한 단백질 ERK1/2와 p38을 유의적으로 활성화 시킨다는 것을 입증 하였다. 이때, ERK1/2 억제제(PD98059)를 처치하여 ERK1/ 2의 활성화가 활성산소종 발생 및 세포사멸에 중요하다는 것을 밝혔다. 따라서, 본 연구 결과는 L-carnitine이 대장 암세포주의 증식을 억제 할 수 있고 이는 대장암의 치료에 있어 잠재적인 치료 물질이 될 수 있음을 시사하며 이 과정에 관여하는 신호전달기전을 조사하여 항암의 치료기 전에서 활성산소종이나 ERK1/2, p38 단백질의 활성화의 중요성을 제시하였다.
It is well known that lymph node metastasis is a major prognostic factor in patients with oral squamous cell carcinoma (OSCC). 30-40% of patients with OSCC have already undergone regional metastasis at diagnosis. The survival rate of patients with metastasis is reduced by more than 50%. Therefore, prevention and early detection of metastasis are very important to increase the survival rate of patients. Many investigators have studied the molecular mechanism of metastasis and tried to develop the molecules to inhibit any step of metastatic cascade. Among those molecules, an interest in the metastasis suppressor gene has been increasing. Expression of metastasis suppressor KiSS-1 has shown to be significantly related to poor clinical outcome and worse survival rate of patient in various malignancies of different organs. In addition, our previous study in OSCC also revealed that downregulation of KiSS-1 expression correlated with the presence of cervical lymph node metastasis, one part of tumor progression. Therefore, further investigation was needed to identify the molecular function of KiSS-1 using OSCC cell line and to evaluate the possibility of KiSS-1 as a new therapeutic target.
Autophagy is recently receiving the spotlight as the development strategy for promising anticancer drugs. In particular, the majority of anticancer drugs originating from natural products are known to induce autophagy. Saururus chinensis has been used for treating various inflammatory diseases. Recent research has revealed that the extract of Saururus chinensis possess cytotoxicity for various types of human cancer cells. However, the exact action mechanism of Saururus chinensis extract for oral squamous cell carcinoma (OSCC) has not been studied yet. Therefore, the authors of this research aim to study the effect of methanol extract of S. chinensis (MESC) on OSCC cells. To observe the cell proliferation inhibitory effect of MESC on HSC3 cells, the authors conducted the trypan blue exclusion assay. Also, the action mechanism of MESC was studied by conducting the cell cycle analysis, acidic vesicular organelle (AVO) staining and flow cytometry analysis, monodansylcadaverine (MDC) staining, propidium iodide staining, and Western blotting on MESC-treated HSC3 cells. When HSC3 cells were treated in MESC, the cell proliferation was suppressed in time-dependent and dose-dependent manners. Also, the number of sub-G1 arrested cells increased in a dose-dependent manner. MDC punctate and AVO puncta significantly increased respectively. Western blot analysis demonstrated the expression of autophagy-related proteins increased, but apoptotic proteins were not observed. Also, the pAkt protein was reduced, while the p-p38 protein and pERK protein increased. According to our results, MESC induced autophagy and accompanied changes in the cell cycle in HSC3 cells. Also, the alteration in Akt, ERK, and p38 pathways were confirmed. This result suggested the possibility of MESC as the new promising adjuvant for treating OSCC patients.
본 연구에서는 발아와 고압처리에 따른 검정콩 조사포닌 추출물의 in-vitro 항암 활성을 확인하기 위하여 검정콩을 발아시킨 후 고압처리하고, 조사포닌 추출물을 제조한 후 인체 유래 유방암(MCF-7), 대장암(HCT-116), 전립선암(PC-3) 및 위암(AGS) 세포주에 대한 증식억제효과를 검토하였다. 대조구의 조사포닌 추출물은 400 μg/mL 농도에서 4종의 암세포주에 대하여 67.02~91.70% 범위의 생존율을 보여 항암 효과가 낮았지만, 발아 4일차 콩을 150 MPa의 압력에서 고압처리한 검정콩의 조사포닌 추출물은 23.94~57.37% 범위의 생존율을 보여 발아와 고압처리에 의해 암세포 성정억제효과가 증가하였다. 특히, 위암세포(AGS)의 경우 다른 세포주에 비해 저농도에서도 세포증식효과를 보여 가장 높은 암세포 증식억제 효과를 나타내었다. 이상의 결과로부터 항암 활성을 나타내는 Soyasaponin, B group의 soyasaponin 및 soyasapogenol의 함량은 발아와 고압처리에 의해 증가하였고, 고압처리 발아콩으로부터 항암 활성을 나타내는 사포닌 추출물의 개발이 가능할 것으로 생각된다. 또한, 추후 항암 활성 물질의 분리동정과 메커니즘 규명에 대한 연구가 수행되어야 할 것으로 판단된다.
Previous studies have shown that kisspeptin (Kp-10) is expressed in mammalian ovaries; however, the expression and role of Kp-10 in bovine ovarian granulosa cells are still unclear. In this study, we assessed the expression of Kp-10 and its effects on the proliferation and apoptosis of bovine granulosa cells. Immunohistochemical analysis showed that Kp-10 was expressed in the cytoplasm of bovine ovarian granulosa cells. Moreover, MTT (3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl-2- H-tetrazolium bromide) assays showed that 100 nM Kp-10 significantly inhibited the viability of granulosa cells (P<0.01). Flow cytometry analysis showed that Kp-10 could significantly increase accumulation of cells in the G1 phase, decrease accumulation of cells in the S phase, and promote apoptosis in bovine granulosa cells (P<0.05). Additionally, Kp-10 decreased the mRNA levels of Bcl-2, an anti-apoptotic gene; increased the mRNA levels of caspase-3, a pro-apoptotic gene; and increased the mRNA levels of Fas and Fasl, two membrane surface molecule genes (P<0.05). Thus, our findings demonstrated for the first time that Kp-10 inhibited proliferation and promoted apoptosis in bovine ovarian granulosa cells. These findings provide insights into our understanding of the role of Kp-10 in mediating the proliferation of bovine granulosa cells.
Induced pluripotent stem cell (iPSC)-derived mesenchymal stem cells (iMSCs) serve as a unique source for cell therapy. We investigated whether exosomes from iMSCs promote the proliferation of human keratinocytes (HaCaT) and human dermal fibroblasts (HDFs). iPSCs were established from human Wharton’s jelly MSCs and were allowed to differentiate into iMSCs. Exosomes were collected from the culture supernatant of MSCs (MSC-exo) and iMSCs (iMSC-exo), and their characteristics were investigated. Both exosome types possessed basic characteristics of exosomes and were taken up by skin cells in vitro and in vivo. A significant increase in HaCaT proliferation was observed with iMSC-exo, although both exosomes increased the viability and cell cycle progression in HaCaT and HDFs. No significant difference was observed in the closure of wound scratch and the expression of reparative genes between cells treated with the two exosome types. Both exosomes enhanced the secretion of collagen in HaCaT and HDFs; however, an increase in fibronectin level was observed only in HaCaT, and this effect was better with iMSC-exo treatment. Only iMSC-exo increased the phosphorylation of extracellular signal-regulated kinase (ERK)-1/2. Our results indicate that iMSC-exo promote the proliferation of skin cells by stimulating ERK1/2 and highlight the application of iMSCs for producing exosomes.
Stem cells have special properties, such as self-renewal, proliferation, and the multilineage differentiation. Generally, stem cells are categorized into embryonic stem cells (ESCs), adult stem cells (ASCs), and induced pluripotent stem cells (iPSCs). Mesenchymal stem cells (MSCs) are a type of ASCs with a multipotent property. MSCs are easily isolated from various tissues and organs in the human body and can differentiation into multiple lineages, such as bone, cartilage, fat, and muscles. Compared to ESCs and iPSCs, MSCs possess less proliferation and differentiation capacities, therefore, a much scientific concern is concerned toward promoting the proliferation and the differentiation potency of MSCs. There are various methods to achieve this goal such as the treatment of various types of small molecules or culturing on specific peptides. Producing of high-quality MSCs with enhanced proliferation and differentiation capacities will definitely be a useful tool for stem cell-mediated tissue regeneration and the further clinical application.
Nicotine of tobacco component has a controversial impact in the clinical outcome of dental implants. Although numerous nicotine effects on bone healing around implants have been presented, it is rarely reported in vitro study about normal human osteoblast(NHost) from oral and maxillofacial area at the surface of implants. The purpose of the present study was to evaluate the effect of nicotine on the proliferation and differentiation response of NHost to plasmatic and salivary levels of nicotine reported in smokers at the surface of screw-type plasma-sprayed titanium implants. NHosts were seeded on the surface of titanium implants and cultured for 21 days in α-MEM supplemented with 10% FBS, 50mg/ml ascorbic acid, 5mM β-glycerophosphate and 100nM dexamethasone. Seeded implants were exposed to various nicotine concentration(0.05-0.5mg/ml) from 1 to 21 days, and characterized for cell morphology, proliferation, differentiation, alkaline phosphatase(ALP) activity and ionized calcium concentration(Cai) of medium. Continuous exposure to higher nicotine concentration(above 0.3mg/ml) induced a dose- and time-dependent vacuolation of the cytoplasm, and a tendency to detach from the implant surface. 0.05mg/ml(lower nicotine concentration) did not cause significant effects in the cell proliferation and ALP activity. 0.1-0.2mg/ml caused evident dose-dependent effects in increased cell proliferation, ALP activity and earlier onset of matrix mineralization at levels up to 0.2mg/ml, while a dose-dependent inhibitory effect at 0.3-0.5mg/ml. Cai concentration of control group was decreased at 14 days. Increased Cai concentration at 0.1-0.2mg/ml, decreased Cai concentration at 0.3mg/ml and no change at 0.5mg/ml during the culture period were seen. It suggested that nicotine concentration could paly an role in modulating NHost activity as a contributing factor associated with proliferation and differentiation of NHost at the surface of implants.
Clover cyst nematode, Heterodera trifolii have recently been identified as a serious pest to Chinese cabbage in highland area of Korea. This study was conducted to investigate the host specificity effect of H. trifolii by their inoculation density and temperature on Chinese cabbage and kale in pot in laboratory condition. H. trifolii inoculated with (1, 2, 4, 8, 16 eggs /g soil), where the number of female resulted higher in Chinese cabbage than in kale but there was no statistical differences between them. The number of cysts did not show any variation by their inoculation density in both plants. The results depicted the proliferation of H. trifolii by temperature (15, 20, 25, 30℃), where the number of females, cysts and eggs has their highest proliferation in 20℃ condition. The study thus demonstrated that, Chinese cabbage is considered a suitable host of the clover cyst nematode compared to the kale and the suitable temperature of the reproduction is estimated as 20°C.
Long-lived Small Modular Reactors are being promoted as an innovative way of catering to emerging markets and isolated regions. They can be operated continuously for decades without requiring additional fuel. A novel configuration of long-lived reactor core employs a mixed neutron spectrum, providing an improvement in nonproliferation metrics and in safety characteristics. Starting with a base sodium reactor design, moderating material is inserted in outer core assemblies to modify the fast spectrum. The assemblies are shuffled once during core lifetime to ensure that every fuel rod is exposed to the thermalized spectrum. The Mixed Spectrum Reactor is able to maintain a core lifetime over two decades while ensuring the plutonium it breeds is below the weapon-grade limit at the fuel discharge. The main drawbacks of the design are higher front-end fuel cycle costs and a 58% increase in core volume, although it is alleviated to some extent by a 48% higher power output.
Micropropagation studies of two Hydrangea. macrophylla cultivars, ‘Blaumeise’ and ‘Vanilla Sky’, were performed by tissue culture. The results showed that supplementing B5 medium with different concentrations of 6-Benzylaminopurine (BA) (0.0, 0.5, 1.0, 1.5, and 2.0 mgL-1) significantly increased the number of shoots and leaves per explant compared with the control in both cultivars. In ‘Blaumeise’, 1.0 mgL-1 of BA was the concentration that achieved the highest number of shoots and leaves, i.e., 12.5 and 93.25, respectively. In ‘Vanilla Sky’ 1.5 mgL-1 of BA resulted in the highest numbers of shoots (4.25) and leaves (29). The longest shoot in both ‘Blaumeise’ (2.10 cm) and ‘Vanilla Sky’ (1.18 cm) was obtained at 0.0 mg·L-1 of BA. The highest number of roots in both ‘Blaumeise’ (11) and ‘Vanilla Sky’ (6.5) were obtained with indole-3-butyric acid (IBA) 1.0 mgL-1 of IBA.