• Asian Pacific Journal of Cancer Prevention
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• 제15권21호
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• pp.9327-9333
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• 2014

Breast cancer is the second most common cancer and second leading cause of cancer deaths in women. Phosphatidylinositol-3-kinase (PI3K)/AKT pathway mutations are associated with cancer and phosphatidylinositol-4, 5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) gene mutations have been observed in 25-45% of breast cancer samples. Insulin growth factor binding protein-5 (IGFBP-5) can show different effects on apoptosis, cell motility and survival in breast cancer. We here aimed to determine the association between PIK3CA gene mutations and IGFBP-5 expressions for the first time in breast cancer patients. Frozen tumor samples from 101 Turkish breast cancer patients were analyzed with high resolution melting (HRM) for PIK3CA mutations (exon 9 and exon 20) and 37 HRM positive tumor samples were analyzed by DNA sequencing, mutations being found in 31. PIK3CA exon 9 mutations (Q546R, E542Q, E545K, E542K and E545D) were found in 10 tumor samples, exon 20 mutations (H1047L, H1047R, T1025T and G1049R) in 21, where only 1 tumor sample had two exon 20 mutations (T1025T and H1047R). Moreover, we detected one sample with both exon 9 (E542Q) and exon 20 (H1047R) mutations. 35% of the tumor samples with high IGFBP-5 mRNA expression and 29.4% of the tumor samples with low IGFBP-5 mRNA expression had PIK3CA mutations (p=0.9924). This is the first study of PIK3CA mutation screening results in Turkish breast cancer population using HRM analysis. This approach appears to be a very effective and reliable screening method for the PIK3CA exon 9 and 20 mutation detection. Further analysis with a greater number of samples is needed to clarify association between PIK3CA gene mutations and IGFBP-5 mRNA expression, and also clinical outcome in breast cancer patients.

• 생명과학회지
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• 제27권6호
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• pp.673-679
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• 2017

미세소관을 따라 이동하는 모터단백질들은 세포내 물질수송에 필수적인 역할을 한다. Kinesin 1은 세포내에서 미세소관을 따라 움직이는 모터단백질로서 다양한 소포, mRNA, 그리고 단백질의 세포내 수송에 관여한다. Kinesin 1은 2개의 장쇄단위체(KHCs, 또는 KIF5s)와 2개의 경쇄단위체(KLCs)로 구성되어 있다. KIF5s는 N-말단에 모터도메인을 가지고 있고 C-말단의 운반체 결합도메인을 통해 다양한 운반체와 결합한다. 본 연구에서 KIF5B와 결합하는 단백질을 분리하기 위하여 효모 two-hybrid 탐색을 수행한 결과 미세소관의 plus end 결합단백질인 cytoplasmic linker protein 170 (CLIP-170)을 분리하였다. CLIP-170의 coiled-coil 도메인은 KIF5B의 운반체 결합도메인과 결합하였다. 또한 CLIP-170은 KIF5A와 KIF5C와도 결합하였다. 그리고 glutathione S-transferase (GST) pull-down을 통해 KIF5s와 CLIP-170이 단백질수준에서 결합함을 확인하였다. 생쥐 뇌파쇄액을 KIF5B 항체로 면역침강한 결과 CLIP-170이 같이 침강함을 확인하였다. 이러한 결과들은 kinesin 1이 세포내에서 CLIP-170을 운반함을 시사한다.

• Advances in Traditional Medicine
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• 제1권1호
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• pp.8-27
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• 2000

Components of extracellular matrix and the matrix-degrading enzymes are some of the key regulators of tumor metastasis and angiogenesis. Hyaluronic acid (HA), a matrix glycosaminoglycan, is known to promote tumor adhesion and migration, and its small fragments are angiogenic. Until now, we have compared levels of hyaluronidase, an enzyme that degrade HA, in normal adult prostate, benign prostate hyperplasia and prostate cancer tissues and in conditioned media from epithelial explant cultures, using a substrate (HA)-gel assay and ELISA-like assay (Kim et al., unpublished results). The present review described an overall characterization of hyaluronidases and its application to human diseases. The hyaluronidases are a family of enzymes that have, until recently, deed thorough explication. The substrate for these enzymes, hyaluronan, is becoming increasingly important, recognized now as a major participant in basic processes such as cell motility, wound healing, embryogenesis, and implicated in cancer progression. And in those lower life forms that torment human beings, hyaluronidase is associated with mechanisms of entry and spread, e.g. as a virulence factor for bacteria, for tissue dissection in gas gangrene, as a means of treponema spread in syphilis, and for penetration of skin and gut by nematode parasites. Hyaluronidase also comprises a component of the venom of a wide variety of organisms, including bees, wasps, hornets, spiders, scorpions, sh, snakes and lizards. Of particular interest is the homology between some of these venom hyaluronidases and the enzyme found in the plasma membrane of mammalian spermatozoa, attesting to the ancient nature of the conserved sequence, a 36% identity in a 300 amino acid stretch of the enzyme protein. Clearly, hyaluronidase is of biological interest, being involved in the pathophysiology of so many important' human disorders. Greater effort should be made in studying this family of enzymes that have, until recently, been overlooked. Also, oriental medical application of the hyaluronidase will be discussed with respect to inhibition and suppression of inflammation and malignacy.

• 한국식품영양과학회지
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• 제42권8호
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• pp.1167-1174
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• 2013

암세포를 포함한 생체 내 빠른 분열을 요구하는 세포 집단에서 세포 내 구성요소 및 에너지원으로서 글루타민의 요구량이 증대되지만, 종양세포의 글루타민 의존적 대사작용에 관한 기전은 여전히 잘 알려진 바 없다. 본 연구에서는 LnCaP 전립선 암세포의 이동성 및 침윤성에 미치는 글루타민 결핍효능을 조사하였다. 본 연구의 결과에 의하면 LnCaP 세포에서 글루타민 결핍에 의하여 세포의 이동성 및 세포의 침윤성이 현저하게 억제되었으며, 이러한 이동성 및 침윤성 억제는 TIMPs의 발현 증대에 의한 MMPs의 발현 감소 및 그들의 효소적 활성 저하와 연관성이 있었다. 또한 글루타민이 결핍된 조건에서 배양된 LnCaP 세포에서 TER의 현저한 증가가 관찰되었는데, 이는 TJs의 조절인자인 claudin family 발현의 차단에 의한 것으로 생각되어진다. 본 연구의 결과에 의하면 암세포의 증식에서 글루타민의 결핍은 TJ의 결합력 증대와 MMPs의 활성을 저하시킴으로써 암세포 전이에 가장 기본적인 과정인 암세포의 이동성과 침윤성을 억제시킬 수 있을 것으로 생각된다.

• 한국식품영양과학회지
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• 제32권6호
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• pp.818-824
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• 2003

전국에 분포되어 있는 우수메주를 일반분석 및 관능평가를 통해 선발하고 선발된 메주의 우수 균주를 분리 동정하여 형태학적 생육특성을 조사하였다. 지역별 메주의 성분분석 결과, 수분함량은 7.2∼28.8%, 조단백질 함량은 32.7∼42.3%의 범위를 나타내었다. 아미노태질소 함량은 경북지역의 메주가 770.8 mg%이었고, 전북지역 메주가 239.9 mg%로 가장 낮았다. 각 지역에서 생산된 메주로 만든 된장의 유리아미노산 함량은 전북 된장이 4,169.6 mg%, glutamic acid는 경북이 499.4 mg%로 가장 높은 함량을 나타내었다. 그리고 메주의 관능검사 결과 경북지역에서 수거한 메주가 색상, 외관, 종합적 품질평가에서 타지역과 유의성(p<0.05) 있게 좋은 것으로 나타났다. 경북지역 메주에서 분리한 균의 형태학적 특성은gram 양성, 호기성 간균이며 운동성이 있었다. 세포의 지방산 조성중 주요 성분의 함량은 $C_{15:0}$ anteiso 지방산이 30.7%, $C_{15:1}$ iso 지방산이 28.9%, $C_{17:0}$ anteiso 지방산이 13.3%,그리고 $C_{17:0}$ iso지방산이 11.2%를 나타내었다. 이 균주의 identification coefficient는 표준균주와 같은 0.653을 나타내었으며, Bergey의 매뉴얼과 지방산 profile에 따라서 분석한 결과 B. licheniformis NH20으로 동정되었다. B. licheniformis NH20의 최적 생육 온도는 32$^{\circ}C$, 최적 초기 pH는 7.0, 최적 염도는 2%, 그리고 배양시간은 9시간이었다. 9시간이었다.

• Korean Journal of Acupuncture
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• 제25권3호
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• pp.147-166
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• 2008

Objectives : Crohn's disease (CD) is characterized by a chronic relapsing inflammation of the bowel in which proinflammatory cytokines play an important perpetuating role. Methods : Mice (preventive animal model of gliotoxin) were treated with 5 % 2,4,6-trinitrobenzenesulfonic acid (TNBS) at day 1 and day 7. To investigate preventive effects of acupuncture with Gujin at $LI_{11}$, acupuncture was carried out at day -1, day 1, day 3. And, to investigate therapeutic effects, acupuncture with Gujin was carried out at day 3, day 5, day 7. For the data analysis, we checked weight and width of colon, diarrhea, edema, survival rate, changes of body weight, and myeloperoxygenase (MPO) activity. For analysing protein expression, we carried out immunohistochemical staining and Western blot and we analyzed mRNA expression by RT-PCR. Results : Colon of TNBS treated mice was erosive and shortening compared with the colon of control mice and induced damages of colon epithelial cell layer and induced infiltration of immune cells in all layer of colon. Acupuncture of gujin at $LI_{11}$ in preventive mode suppressed macorscopic damages such as erosive and shortening of colon by TNBS and damages of intestinal epithelial cells and infiltration of immune cells in the colon. The average weight of 5 cm distal colon was increased in TNBS treated mice (758${\mu}g$) compared with in control mice (112${\mu}g$) and width of distal colon was also increased in TNBS treated mice (4.9mm) compared with in control mice (1.3mm). Acupuncture with Gujin at $LI_{11}$ in preventive and therapeutic mode suppressed increase of colon weight and width by TNBS. TNBS induced edema of colon and diarrhea and Acupunctured with Gujin at $LI_{11}$ in preventive and therapeutic mode ameliorated these symptom by TNBS. In preventive and therapeutic mode, the effects of acupuncture with Gujin at $LI_{11}$ were increasing the motility, suppressing body weight decreasing, suppressing MPO activity, reducing expressing of TNF-${\alpha}$, IL-1b, and ICAM-1 in colon compared with that by TNBS Conclusions : This study demonstrates that acupuncture with Gujin at $LI_{11}$ represents a potential therapeutic method of Crohn's disease.

• The Korean Journal of Physiology and Pharmacology
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• 제15권3호
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• pp.129-135
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• 2011

In this study we determined whether or not 5-hydroxytryptamine (5-HT) has an effect on the pacemaker activities of interstitial cells of Cajal (ICC) from the mouse small intestine. The actions of 5-HT on pacemaker activities were investigated using a whole-cell patch-clamp technique, intracellular $Ca^{2+}$ ($[Ca^{2+}]_i$) analysis, and RT-PCR in ICC. Exogenously-treated 5-HT showed tonic inward currents on pacemaker currents in ICC under the voltage-clamp mode in a dose-dependent manner. Based on RT-PCR results, we found the existence of 5-$HT_{2B,\;3,\;4,\;and\;7}$ receptors in ICC. However, SDZ 205557 (a 5-$HT_4$ receptor antagonist), SB 269970 (a 5-$HT_7$ receptor antagonist), 3-tropanylindole - 3 - carboxylate methiodide (3-TCM; a 5-$HT_3$ antagonist) blocked the 5-HT-induced action on pacemaker activity, but not SB 204741 (a 5-$HT_{2B}$ receptor antagonist). Based on $[Ca^{2+}]_i$ analysis, we found that 5-HT increased the intensity of $[Ca^{2+}]_i$. The treatment of PD 98059 or JNK II inhibitor blocked the 5-HT-induced action on pacemaker activity of ICC, but not SB 203580. In summary, these results suggest that 5-HT can modulate pacemaker activity through 5-$HT_{3,\;4,\;and\;7}$ receptors via $[Ca^{2+}]_i$ mobilization and regulation of mitogen-activated protein kinases.

• 한국식품영양과학회지
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• 제43권1호
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• pp.86-92
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• 2014

Cordycepin은 C. militaris의 주요 생리활성 물질로서 인체 면역기능 강화, 항염증, 항산화, 항노화 및 항암활성을 포함한 다양한 약리효능이 있는 것으로 알려져 있다. 본 연구에서는 HCT116 대장암세포를 이용하여 암전이의 주요 과정인 암세포의 이동성 및 침윤성에 미치는 cordycepin의 효능에 관하여 조사하였다. 본 연구의 결과에 의하면 세포독성이 없는 범위에서 cordycepin은 HCT116 세포의 이동성과 침윤성을 유의적으로 억제하였다. RT-PCR 및 Western blotting 결과에 의하면 cordycepin은 TJs의 주요 구성인자인 claudin family 인자들의 발현을 억제하였으며, 이는 TJ의 전기적 저항성의 증대와 연관이 있었다. Cordycepin은 또한 MMP-2 및 -9의 발현과 활성을 저해함과 동시에 TIMP-1 및 -2의 발현은 증가시켰다. 따라서 cordycepin에 의한 HCT116 대장암세포의 전이능 억제는 TJ의 견고성 증대와 MMPs의 활성 억제와 연관성이 있음을 알 수 있었다.

• 대한미생물학회지
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• 제22권3호
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• pp.209-218
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• 1987

Leptospira interrogans, the causative organism of leptospirosis, is characterized by a fine helical morphology, and the helix is almost always right-handed. However, one of the striking features of recent isolates of L. interrogans in Korea was the heterogeneity in their morphology. Even under optimal culture conditions($30^{\circ}C$, EMJH medium), rods, spiral forms with right or left-handed helices, and even spherical forms of L. interrogans were present. Although the literature notes the presence of left-handed helices, long rods, and spherical forms in cultures of L. interrogans isolates, little is known about the cause of this morphologic heterogeneity. In an attempt to answer this question, this study was initiated to examine the effects of culture conditions, especially temperature and medium, on the morphology of L. interrogans. Four temperatures($5^{\circ}C$, $15^{\circ}C$, $30^{\circ}C$, and $37^{\circ}C$) and two types of media(Fletcher and EMJH) were used; one strain from Korean isolates and L. interrogans serovar canicola obtained from the Pasteur Institute(Paris, France) were employed throughout the study. The findings are as follows: 1. The L. interrogans isolated in Korea(UM-19) had a larger cell diameter($0.25{\sim}0.30\;{\mu}m$: $0.10{\sim}0.15\;{\mu}m$), and helix diameter($0.10{\sim}0.60\;{\mu}m$: $0.10{\sim}0.15\;{\mu}m$) than that obtained from the Pasteur Institute, but they varied in their distances between the helices($0.31{\sim}1.00\;{\mu}m$: $0.50{\sim}0.70\;{\mu}m$). 2, When UM-19 was grown at $37^{\circ}C$ after months or longer preincubation at $5^{\circ}C$ or $15^{\circ}C$, the majority of the organisms were spiral forms; however, they became rods when subcultured at $30^{\circ}C$ or $37^{\circ}C$. No significant morphological differences were found between Fletcher and EMJH media. 3. When L. interrogans serovar canicola was subcultured more than ten times at $37^{\circ}C$, some of the organism lost their motility as well as the hooks at either one or both ends, but only in Fletcher medium. The number of variants increased with the frequency of subculturing. These findings suggested that L. interrogans strain (UM-19) is different, in their morphology, from that of the Pasteur Institute, and its various morphologies may represent stages of the life cycle and vary with incubation temperature.

• 생명과학회지
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• 제29권9호
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• pp.1010-1015
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• 2019

카할세포는 위장관에서 향도잡이 역할을 한다. 본연구에서는 생쥐 소장 카할세포에서 발생되는 향도잡이 기능에서 올란자핀의 역할을 연구하였다. 패치클램프 방법을 사용하여 향도잡이 전압을 측정하였다. 올란자핀에 의해서 카할세포 향도잡이 전압이 탈분극 되었으며, 이 탈분극은 무스카린성 3번 수용체 억제제에 의해서 억제 되었다. 세포내 $GDP{\beta}S$을 넣어주니 올란자핀에 의해 향도잡이 전압 탈분극이 억제되었다. 또한, 세포밖 $Na^+$ 농도 감소와 비선택성 양이온 통로 억제제에 의해서 올란자핀에 의한 향도잡이 전압 탈분극이 억제 되었다. 세포내 PLC기전의 억제제인 U-73122에 의해서 올란자핀에 의한 향도잡이 전압 탈분극이 억제 되었다. 이러한 결과로 올란자핀은 무스카린성 3번 수용체를 통해서 세포내 G 단백질과 PLC기전 및 세포밖 $Na^+$이 관여함을 알 수 있었다. 따라서 올란자핀은 카할세포를 통해서 장운동성을 조절 할 수 있을 것으로 생각된다.