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Impact of HER2 and PTEN Simultaneous Deregulation in Non-small Cell Lung Carcinoma: Correlation with Biological Behavior

  • Panagiotou, Ioannis;Georgiannos, Stavros N.;Tsiambas, Evangelos;Karameris, Andreas;Konstantinou, Marios;Lazaris, Andreas C.;Kavantzas, Nikolaos;Vilaras, George;Patsouris, Efstratios
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.6311-6318
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    • 2012
  • Background: HER2/neu overexpression due to gene amplification is an important factor in breast cancer, modifying the sensitivity to anti-HER2 monoclonal antibody therapy. The clinical significance of HER2 expression in non small cell lung carcinoma (NSCLC) is currently under evaluation. The tumor suppressor gene PTEN negatively regulates the HER2/PI3K/Akt signalling pathway. The purpose of this study was to evaluate the role of simultaneous alteration in HER2 and PTEN protein expression in relation to biological behaviour of NSCLCs. Materials and Methods: Protein expression was determined by immunohistochemistry in sixty-one (n=61) NSCLC cases along with CISH for HER2 gene analysis and detection of chromosome 17 aneuploidy. Patients were followed-up for a period of 34 to 41 months after surgery. Results: HER2 overexpression (2+/3+score) was detected in 17 (27.9%) patients while loss of PTEN expression was observed in 24 (39.3%) cases, low expression in 29 (47.6%) and overexpression in 8 (13.1%). Simultaneous HER2 overexpression and PTEN low/loss of expression were correlated with metastasis (71.4% vs 36.2% p=0.03). Analysis in the subgroup of 22 patients of pTNM stage III with lymph node status N1 or N2 revealed that there was a relationship between the number of positive regional lymph node groups and simultaneous deregulation of the two genes (p=0.04). Multivariate analysis determined that HER2 overexpression was associated with an increasing risk of developing metastases (OR: 4.3; 95%CI: 1.2-15.9; p: 0.03) while PTEN overexpression was associated with lower risk (OR: 0.1; 95%CI: 0.1, 1.0; p: 0.05). Conclusions: Simultaneous HER2/PTEN deregulation is a significant genetic event that leads to a more aggressive phenotype of NSCLC.

Expression and Clinical Significance of mTOR in Surgically Resected Non-small Cell Lung Cancer Tissues: a Case Control Study

  • Liu, Zhe;Wang, Liang;Zhang, Li-Na;Wang, Yue;Yue, Wen-Tao;Li, Qi
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.6139-6144
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    • 2012
  • Aims: Mammalian target of rapamycin (mTOR) is master regulator of the PI3K/Akt/mTOR pathway and plays an important role in NSCLCs. Here we characterized mRNA and protein expression levels of mTOR and its functional associated molecules including PTEN, IGF-1R and 4EBP1 in surgically resected NSCLCs. Methods: Fifty-four patients with NSCLCs who underwent pulmonary resection were included in current study. mRNA levels of mTOR, PTEN, IGF-1R, and 4EBP1 were evaluated by RT-PCR and protein expression of mTOR, PTEN, and IGF-1R by immunohistochemistry (IHC). Association of expression of the relevant molecules with clinical characteristics, as well as correlations between mTOR and PTEN, 4EBP1 and IGF-1R were also assessed. Results: The results of RT-PCR showed that in NSCLCs, the expression level of mTOR increased, while PTEN, 4EBP1 and IGF-1R decreased. Statistical analysis indicated high IGF-1R expression was correlated with advanced clinical stage (stage III) and PTEN expression was reversely associated with tumor size (P=0.16). The results of IHC showed mTOR positive staining in 51.8% of cases, while IGF-1R positive staining was found in 83.3% and loss of PTEN in 46.3%. Protein expression of mTOR was correlated with its regulators, PTEN and IGF-1R, to some extent. Conclusions: Abnormal activation of mTOR signaling, high expression of IGF-1R, and loss of PTEN were observed in resected NSCLC specimens. The poor expression agreement of mTOR with its regulators, PTEN, and IGF-1R, implied that combination strategy of mTOR inhibitors with other targets hold significant potential for NSCLC treatment.

Experimental Study on Inhibition Effects of the XAF1 Gene against Lung Cancer Cell Proliferation

  • Yang, Wen-Tao;Chen, Dong-Lai;Zhang, Fu-Quan;Xia, Ying-Chen;Zhu, Rong-Ying;Zhou, Duan-Shan;Chen, Yong-Bing
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.18
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    • pp.7825-7829
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    • 2014
  • Objective: To investigate the effect of high expression of XAF1 in vivo or in vitro on lung cancer cell growth and apoptosis. Methods: 1. The A549 human lung cancer cell line was transfected with Ad5/F35 - XAF1, or Ad5/F35 - Null at the same multiplicity of infection (MOI); (hereinafter referred to as transient transfected cell strain); XAF1 gene mRNA and protein expression was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting respectively. 2. Methyl thiazolyl tetrazolium (MTT) and annexin V-FITC/PI double staining were used to detect cell proliferation and apoptosis before and after infection of Ad5/F35 - XAF1 with Western blotting for apoptosis related proteins, caspase 3, caspase - 8 and PARP. 3. After the XAF1 gene was transfected into lung cancer A549 cells by lentiviral vectors, and selected by screening with Blasticidin, reverse transcription polymerase chain reaction (RT-PCR) and Western blotting were applied to detect mRNA and protein expression, to establish a line with a stable high expression of XAF1 (hereinafter referred to as stable expression cell strain). Twenty nude mice were randomly divided into groups A and B, 10 in each group: A549/XAF1 stable expression cell strain was subcutaneously injected in group A, and A549/Ctrl stable cell line stable expression cell strain in group B (control group), to observe transplanted tumor growth in nude mice. Results: The mRNA and protein expression of XAF1 in A549 cells transfected by Ad5/F35 - XAF1 was significantly higher than in the control group. XAF1 mediated by adenovirus vector demonstrated a dose dependent inhibition of lung cancer cell proliferation and induction of apoptosis. This was accompanied by cleavage of caspase -3, -8, -9 and PARP, suggesting activation of intrinsic or extrinsic apoptotic pathways. A cell strain of lung cancer highly expressing XAF1 was established, and this demonstrated delayed tumor growth after transplantation in vivo. Conclusion: Adenovirus mediated XAF1 gene expression could inhibit proliferation and induce apoptosis in lung cancer cells in vitro; highly stable expression of XAF1 could also significantly inhibit the growth of transplanted tumors in nude mouse, with no obvious adverse reactions observed. Therefore, the XAF1 gene could become a new target for lung cancer treatment.

Berchemia floribunda-mediated Proteasomal Degradation of CyclinD1 via GKS3β-dependent Threonine-286 Phosphorylation in Human Colorectal Cancer Cells (인간 대장암 세포에 대한 먹넌출 추출물의 GSK3β 의존성 threonine-286 인산화를 통한 Cyclin D1 분해)

  • Kang, Yeongyeong;Eo, Hyun Ji;Kim, Da Som;Park, Youngki;Song, Jeong Ho;Park, Gwang Hun
    • Korean Journal of Plant Resources
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    • v.33 no.4
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    • pp.271-278
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    • 2020
  • In this study, we evaluated the anti-cancer activity and potential molecular mechanism of 70% ethanol extracts of the Berchemia floribunda (BF) which belongs to Rhamnaceae against human colorectal cancer cells. The treatment of BF decreased the cell proliferation in HCT116 cell and suppressed cellular accumulation of Cyclin D1 protein. Inhibition of proteasomal activity by MG132 attenuated BF-mediated Cyclin D1 downregulation and Cyclin D1 was decreased in the cell treated with BF. These findings indicates that BF-mediated Cyclin D1 downregulation may be result from Cyclin D1 proteasomal degradation. Additionally, BF-mediated Cyclin D1 degradation was blocked in the presence of LiCl, a GSK3β inhibitor, but not PD98059, SP600125, SB203580, Bay11-7082, LY294002 an ERK1/2 inhibitor, JNK inhibitor, p38 inhibitor, IκK inhibitor and PI3K inhibitor. Furthermore, BF phosphorylated Cyclin D1 at threonine-286 (Thr286), and LiCl-induced GSK3β inhibition reduced the BF mediated phosphorylation of Cyclin D1 at Thr286. These results suggested that BF may downregulate Cyclin D1 expression as a potential anti-cancer target through GSK3β dependent Cyclin D1 degradation. Therefore, this study provides that the extract of BF has anticancer activity against human colorectal cancer cells.

Synthesis of New 4-Oxo-2-Thioxo-1,2,3,4-Tetrahydropyrimidine Derivatives with an Incorporated Thiazolidinone Moiety and Testing Their Possible Serine Protease and Cercarial Elastase Inhibitory Effects with a Possible Prospective to Block Penetration of Schistosoma mansoni Cercariae into the Mice Skin

  • Bahgat Mahmoud Mohamed;Maghraby Amany Sayed;Heiba Mogeda Emam;Ruppel Andreas;Fathalla Omar Abd-elfattah Mohamed
    • Archives of Pharmacal Research
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    • v.28 no.9
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    • pp.1002-1012
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    • 2005
  • 5-Substituted 4-oxo-2-thioxo-1,,2,3,4-tetrahydropyrimidine were synthesized by interaction of 4­oxo-2-thioxo-1,2,3,4-tetrahydropyrimidine-5-sulfonylhydrazide with some aldehydes to give the corresponding Schiff-bases, which after cyclization gave corresponding thiazolidinones. For some of the thiazolidinones, Mannich bases reaction was carried out. All the derivatives were tested for their possible inhibitory effect on Schistosoma mansoni cercarial elastase (CE). Only, N'-(4-methylbenzyledine)-4-oxo-2-thioxo-1,2 ,3,4-tetrahydropyrimidine-5-sulfonylhydrazide was found to have potent inhibitory effect on the CE activity with $IC_{50} = 264{\mu}M.$ Upon its use as a paint for mice tails before infection with S. mansoni cercariae, the compound formulated in jojoba oil caused a significant reduction ($93\%$; P-value = 0.0002) in the worm burden. IgG & IgM in mice sera were measured by using several S. mansoni antigens by ELISA. Sera from treated infected mice (TIM) 2, 4, and 6 weeks (W) post infection (PI) showed 1.2 folds lower, 1.2 folds higher, 1.7 folds lower IgM reactivity against soluble cercarial antigenic preparation (CAP), respectively, when compared with sera collected from infected untreated mice (IUM). Sera from TIM 2, 4, and 6WPI showed 1.3, 1.6, and 1.7 folds higher IgG reactivity, respectively against CAP than the IgG reactivity from IUM. Sera from TIM 2, 4 and 6WPI showed 1.5, 1.2 folds lower and 1.4 folds higher IgM reactivity, respectively against soluble worm antigenic preparation (SWAP) when compared with sera collected from IUM. Sera from TIM 2, 4, and 6WPI showed 1.4, 1 folds lower and 1 fold higher IgG reactivity, respectivley to SWAP when compared with sera from IUM. Sera from TIM 2, 4, and 6WPI had generaly lower IgM and IgG reactivities against soluble egg antigen (SEA) when compared with sera from IUM.

A Study on the Daily Probability of Rainfall in the Taegu Area according to the Theory of Probaility (대구지방(大邱地方)의 확률일우량(確率日雨量)에 관(關)한 연구(硏究))

  • Kim, Young Ki;Na, In Yup
    • Economic and Environmental Geology
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    • v.4 no.4
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    • pp.225-234
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    • 1971
  • With the advance of civilization and steadily increasing population rivalry and competition for the use of the sewage, culverts, farm irrigation and control of various types of flood discharge have developed and will be come more and more keen in the future. The author has tried to calculated a formula that could adjust these conflicts and bring about proper solutions for many problems arising in connection with these conditions. The purpose of this study is to find out effective sewage, culvert, drainage, farm irrigation, flood discharge and other engineering needs in the Taegu area. If demands expand further a new formula will have to be calculated. For the above the author estimated methods of control for the probable expected rainfall using a formula based on data collected over a long period of time. The formula is determined on the basis of the maximum daily rainfall data from 1921 to 1971 in the Taegu area. 1. Iwai methods shows a highly significant correlation among the variations of Hazen, Thomas, Gumbel methods and logarithmic normal distribution. 2. This study obtained the following major formula: ${\log}(x-2.6)=0.241{\xi}+1.92049{\cdots}{\cdots}$(I.M) by using the relation $F(x)=\frac{1}{\sqrt{\pi}}{\int}_{-{\infty}}^{\xi}e^{-{\xi}^2}d{\xi}$. ${\xi}=a{\log}_{10}\(\frac{x+b}{x_0+b}\)$ ($-b<x<{\infty}$) ${\log}(x_0+b)=2.0448$ $\frac{1}{a}=\sqrt{\frac{2N}{N-1}}S_x=0.1954$. $b=\frac{1}{m}\sum\limits_{i=1}^{m}b_s=-2.6$ $S_x=\sqrt{\frac{1}{N}\sum\limits^N_{i=1}\{{\log}(x_i+b)\}^2-\{{\log}(x_0+b)\}^2}=0.169$ This formule may be advantageously applicable to the estimation of flood discharge, sewage, culverts and drainage in the Taegu area. Notation for general terms has been denoted by the following. Other notations for general terms was used as needed. $W_{(x)}$ : probability of occurranec, $W_{(x)}=\int_{x}^{\infty}f_{(n)}dx$ $S_{(x)}$ : probability of noneoccurrance. $S_{(x)}=\int_{-\infty}^{x}f_(x)dx=1-W_{(x)}$ T : Return period $T=\frac{1}{nW_{(x)}}$ or $T=\frac{1}{nS_{(x)}}$ $W_n$ : Hazen plot $W_n=\frac{2n-1}{2N}$ $F_n=1-W_x=1-\(\frac{2n-1}{2N}\)$ n : Number of observation (annual maximum series) P : Probability $P=\frac{N!}{{t!}(N-t)}F{_i}^{N-t}(1-F_i)^t$ $F_n$ : Thomas plot $F_n=\(1-\frac{n}{N+1}\)$ N : Total number of sample size $X_l$ : $X_s$ : maximum, minumum value of total number of sample size.

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Production and Consumption of Goat Milk Products in Korea (한국의 산양유제품 생산 및 소비 현황)

  • Park, Seung-Yong
    • Journal of Dairy Science and Biotechnology
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    • v.24 no.2
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    • pp.39-45
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    • 2006
  • There has been a successful re-entry in the form of infant foods and as a product concept of "well-being" milk by feeding goat natural medicinal plants in high mountain lands and goats eating natural feeds. Typical composition of cow's milk and goat's milk are not significantly differ in major nutritional constituents. However, the noticeable differences between milks of the bovine and caprine species concern in the dimensions of the micelles, in casein composition, in size of the micelles and in the mineral charge of the micelle, but the ratio Ca/Pi in the micelle is very close for the two species The potential market in Korea could be expected to expand by keeping its freshness and nutritional benefits. The supply of goat milk products all year around is also an important to the consumers. In order to increase its market scale of goat milk, product manufacturers need extensive advertising promotion. Domestically, goat milk is currently manufactured at small scale dairy goat milk companies and consumed mainly in the form of fresh or fermented goat milk, while imported goat milk powder is used to produce infant goat milk formula by major dairy companies. Decreasing the unpleasant goaty flavour for the Korean consumers would be essential for the researchers who work for dairy science and technology.

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Assessment of DNA Ploidy Patterns in Connection with Prognostic Factors in Patients with Papillary Thyroid Carcinoma (유두상 갑상선암에서 예후인자와 DNA배수성의 상관관계)

  • Chung Woung-Youn;Lee Jong-Hoon;Park Cheong-Soo
    • Korean Journal of Head & Neck Oncology
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    • v.12 no.2
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    • pp.153-160
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    • 1996
  • Regardless of the prognostic factors in papillary thyroid cancer, such as sex, age, size of tumor, extent of disease, and distant metastasis, the prognosis of papillary thyroid cancer is sometimes difficult to predict from clinical and microscopic analysis alone and additional prognostic indicators are needed. Recent studies of thyroid cancer have indicated that DNA aneuploidy may be correlated to the biological behavior of malignancy and inversely correlated to the prognosis, but it still remains contraversal. We performed this study to assess DNA ploidy patterns in relation with the previously known prognostic factors in AMES scoring system and lateral neck node metastasis in papillary thyroid cancer. A series of 132 patients with papillary thyroid cancer and 80 patients with benign thyroid tumor(27 follicular adenomas and 53 adenomatous goiters) as a control group from October 1993 to Feburary 1995 were analyzed and their nuclear DNA content was measured with flow cytometry using fresh tissue specimens. DNA aneuploidy was found in 8(6.1%) in papillary cancer and 8(10%) in benign tumor. S-phase traction(SFP) and proliferative index(PI) were higher in thyroid cancers, being 2.18$\pm$4.24%, 6.34$\pm$4.94% in the papillary thyroid cancers and 1.97$\pm$2.93%, 4.44$\pm$3.80% in the benign tumors, respectively. However there was no significant difference of values between two groups(p>0.05). Among variable prognostic factors studied(age, sex, size of tun or, extent of disease, distant metastasis in AMES scoring system and lateral neck node metastasis), DNA aneuploidy was found to be common in distant metastasis(p<0.001) and in lateral neck node metastasis(p>0.035), but there was no significant difference between the high risk and low risk group according to the AMES scoring system(p<0.08). In our study, DNA aneuploidy was not valuable in determining the presence of malignancy and did not correlate to the AMES scoring system. However, follow-up study of more cases will be needed for accurate information about the DNA ploidy as a independent prognostic factor.

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Cleavage of In Vitro Fertilized Oocytes and Viability of Sperm Cryopreserved with L-Cysteine in Korea native cattle (L-Cysteine을 첨가하여 동결-융해한 한우 정자의 생존성과 체외 수정 난자의 분할)

  • Park, Bola;Lee, Kung-Jin;Lee, Sang-Hee;Lee, Eunsong;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Journal of Embryo Transfer
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    • v.28 no.3
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    • pp.193-198
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    • 2013
  • This study was designed to evaluate the effect of L-cysteine on sperm characteristics and oocyte cleavage in vitro in Korean native cattle. For this study, the freezing of diluted semen were added with Triladyl containing 20% egg-yolk and/or 0, 5, 10 and 20 mM L-cysteine before cryopreservation. The viability in frozen-thawed sperm were estimated by SYBR14/PI double stain, acrosome damage with FITC-PNA, mitochondria intact with Rhodamin123 and hydrogen peroxide($H_2O_2$) level with carboxy-DCFDA by flow-cytometry. The developmental capacity was also assessed with cleavage rates in oocytes fertilized in vitro by frozen-thawed sperm. In results, the sperm viability was significantly increased in 10 mM and 20 mM concentrations of L-cysteine than other groups (p<0.05). In addition, acrosome damage was significantly decreased in 10 mM and 20 mM concentrations of L-cysteine than other groups (p<0.05). The mitochondria intact was also significantly increased in 10 mM and 20 mM concentrations of L-cysteine than other groups (p<0.05). On the other hand, the cleavage rates were significantly increased in 0 mM, 5 mM and 10 mM groups than 20 mM concentration of L-cysteine (p<0.05). The oocyte degeneration of oocytes were significantly decreased in 0 mM, 5 mM and 10 mM groups than in 20 mM L-cysteine group (P<0.05). However, there are no significantly differences among the L-cysteine treatment groups. We suggest that concentration of 10 mM L-cysteine have beneficial impact for sperm cryopreserved in Korean native cattle. This result also could be recommended for artificial insemination program if supported by an improvement in the fertility results and required further study.

Control Efficacy of Brassicaceae Cover Crops against Clover Cyst Nematode, Heterodera trifolii (배추과 녹비작물의 클로버씨스트선충 방제 효과)

  • Ko, Hyoung-Rai;Kim, Se-Jong;Lee, Jae-Kook
    • Research in Plant Disease
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    • v.26 no.2
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    • pp.116-119
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    • 2020
  • To investigate the effects of Brassicaceae cover crops on clover cyst nematode, Heterodera trifolii, 10 cultivars consisting of six of oil radish and four of white mustard were planted in the nematode infected field at Jeongseon city. Two months after planting, the cover crops were plow down and incorporated into the soil using rotavator, decomposed for 1 month, then transplanted kimchi-cabbages. After 70 days, the density of eggs inside of the cyst and the number of females in the soil were examined. As a result, the reproduction rates of eggs in each plots of Adios and Anaconda cultivars, which were 0.04 and 0.02, respectively, were greatly reduced. The number of females in the plots of above two cultivars showed means at 2.5 and 3.5 per 300 ㎤ soil, which were lower than those of other plots. In addition, fresh weights of three plants in the two plots, which were 7.67 and 7.35 kg, were significantly higher than that of the control plot. Collectively, these results suggest that the two cultivars of Brassicaceae cover crops, Adios and Anaconda, could be used for reducing the cyst nematode density.