• Asian Pacific Journal of Cancer Prevention
• /
• 제13권5호
• /
• pp.2235-2240
• /
• 2012

c-Src is one member of non-receptor tyrosine kinase protein family that has over expression and activation in many human cancer cells. It has been shown that c-Src is implicated in various downstream signaling pathways associated with EGFR-dependent signaling such as MAPK and STAT5 pathways. Transactivation of EGFR by c-Src is more effective than EGFR ligands. To inhibit the c-Src expression, we used c-Src antisense oligonucleotide complexed with PAMAM Denderimes. The effect of c-Src antisense oligonucleotide on HT29 cell proliferation was determined by MTT assay. Then, the expression of c-Src, EGFR and the genes related to EGFR-depended signaling with P53 was applied by real time PCR. We used western blot analysis to elucidate the effect of antisense on the level of c-Src protein expression. The results showed, c-Src antisense complexed with PAMAM denderimers has an effective role in decrease of c-Src expression and EGFR-dependent downstream genes.

• 한국발생생물학회:학술대회논문집
• /
• 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
• /
• pp.66-66
• /
• 2003

Protein tyrosine kinases는 표적단백질의 tyrosine 잔기를 인산화하는 효소로서 다양한 종류의 성장인자, peptide 호르몬, cytokine 수용체 하위의 세포 내 신호전달에 관여한다. Non-receptor tyrosine kinase의 일종인 c-Src는 세포막에서 발생한 ligand-receptor 상호작용 하위의 신호전달에서 중요한 역할을 하며 C-terminal Src kinase (Csk)는 Src kinase의 C-terminal tyrosine 잔기를 인산화시켜 Src kinase의 활성을 저해한다. 이러한 Src-Csk loop를 통한 세포 내 신호전달과정은 세포의 증식과 분화, 사멸 조절에 중요한 기능을 갖지만 정소의 발생과 분화 과정에서 Src-Csk loop의 발현 및 정자형성 과정에서의 기능은 밝혀지지 않았다. 본 연구에서는 생쥐 정소에서 출생 후 성적 성숙과정에서 Csk의 발현과 Src kinase 활성의 변동을 조사하였다. Csk mRNA 발현은 생 후 2주령 이하의 미성숙 정소에서 다량으로 발현되었고 사춘기 정소 이후에는 오히려 감소하였다. Csk 단백질의 발현 양상은 mRNA 발현양상과 일치하였다. c-Src kinase 활성은 생 후 2주에 급격히 증가하고 이 후 4주령에서 감소하다가 성체 (8주령)에서 다시 증가하여 가장 높았다. 성체 조직의 Csk 단백질 현존량이 미성숙 개체보다 적은 반면 Src kinase 활성은 가장 높아 Csk 발현의 감소는 Src kinase 활성을 증가하는 것으로 사료된다. 면역조직화학방법으로 정소 조직 내 Csk의 발현양상을 조사한 결과 Leydig cell, Sertoli cell, germ cell 등 도처에서 발현되었으며 Sertoli cell 에서의 발현은 세정관 상피의 구성에 따른 차이가 확인되었다. 성체의 세정관 내에서는 감수분열 이후의 정세포(spermatid)를 감싸고 있는 Sertoli cell의 강소측에서 강한 Csk 활성이 검출되어 생식세포의 분화과정 동안 세정관 상피의 조직재구성에 관여하는 것으로 사료된다. Leydig cell에서의 발현은 생후 1주령까지는 미미하였으나 이후 2주령 이후에는 다량으로 발현함이 확인되어 adult type Leydig cell에서 진행되는 steroidogenesis와의 관련성을 추측할 수 있다. 미성숙 정소로부터 분리한 Sertoli cell-enriched culture에 200 nM testosterone을 처리하였을 때 Csk mRNA의 발현의 증가를 확인할 수 있었으므로 androgen에 의한 Sertoli cell의 분화과정에 Csk가 관여하고 있음을 알 수 있다. 결론적으로 성적 성숙에 따른 생쥐 정소 내 Src-Csk loop의 발현과 Src kinase 활성의 변동은 정소 내 간충조직, 세정관 상피의 증식 및 기능적 분화 과정을 매개하는 생리적 활성분자 수용체 하위의 신호전달 과정에 Src-Csk loop에 의한 조절가능성을 확인할 수 있었다.

• 생명과학회지
• /
• 제17권3호통권83호
• /
• pp.350-355
• /
• 2007

흰쥐유래의 비만세포인 RBL-2H3 세포는 다양한 Src-family kinase를 발현한다. 현재까지의 연구결과에 의하면 비만세포의 초기 활성화에 Lyn kinase가 중요한 역할을 한다고 알려져 왔다. 그러나 그 세포에서 발현되는 다양한 다른 Src-family kinase의 역할은 불분명하다. 본 연구에서는 비만세포에서 다양한 Src-family kinase가 세포내 다른 곳에서 다양하게 발현되고 있다는 사실을 RT-PCR, immunoblotting 그리고 confocal microscopy 기법을 이용하여 증명하였다. 그 결과 Lyn 및 Fgr kinase는 세포막에 위치하고 c-Src 및 Yes kinase는 세포 내 과립에 존재하는 것을 알 수 있었다. 모든 Src-family kinase를 클로닝하고 과발현하여 탈과립 대한 영향을 평가하였다. 그 결과 fyn과 Fgr kinase는 비만세포에서 항원 유도의 탈과립을 증가시켰으며 반면 Lyn kinsae는 탈과립을 억제시키는 것을 확인할 수 있었다. 이러한 결과는 비만세포 초기 신호전달계에서 Fgr가 중요한 역할을 할 가능성을 제시한다.

• Journal of Ginseng Research
• /
• 제22권2호
• /
• pp.133-139
• /
• 1998

We have studied an activation mechanism of $pp60^{c-src}$ protein tyroslne kinase (PTK) by ginsenoside-$Rg_1$ (G-$Rg_1$ ) in NIH(pMcsrc/foc)B c-src overexpressor cells. It was previously reported that G--$Rg_1$ stimulated the activation of c-src kinase at 20 pM with a 18 hr-incubation, increasing the activity by 2-4-fold over that of untreated control, and this effect was blocked by treatments of in- hibitors of either protein synthesis (cycloheximide) or RNA synthesis (actinomycin D) (Hong, H.Y. et at. Arch. Pharm. Res. 16, 114 (1993)). However, an amount of c-src protein itself in wild-type cells was not changed by G-$Rg_1$. When the cells mutated at one or two tyrosine residue(s) (Y416/527) that are important sites to regulate the kinase activity were treated with G-$Rg_1$, increases both in the activity of c-src kinase and in the expression of the protein were not observed. In addition, removal of extracellular calcium ion by EGTA or inhibition of PKC by H-7 canceled the G-$Rg_1$-induced activation of the kinase. Although the activation was little affected by G-$Rg_1$ with a calcium ionophore A23187, it was synergistically stimulated by treatment of G-Rgl and PMA, a PKC activator. Taken together, these results suggest that the activation of c-src kinase by G-$Rg_1$ is caused by an increase in the specific activity of the kinase, but not in amount of it, and is involved with both collular calcium ion and PKC. Further the increase in the specific activity of c-src kinase may result from altered phosphorylation at tyro-416 and -527.

• Asian Pacific Journal of Cancer Prevention
• /
• 제13권9호
• /
• pp.4751-4756
• /
• 2012

Colon cancer continues to be one of the most common cancers, and the importance and necessity of new therapies needs to be stressed. The most important proto-oncogen factors for colon cancer appear to be epidermal growth factor receptor, EGFR, and c-Src with high expression and activity leading to tumor growth and ultimately to colon cancer progression. Application of c-Src and EGFR antisense agents simultaneously should theoretically therefore have major benefit. In the present study, anti-EGFR and c-Src specific antisense oligodeoxynucleotides were combined in a formulation using PAMAM dendrimers as a carrier. Nano drug entry into cells was confirmed by flow cytometry and fluorescence microscopy imaging and real time PCR showed gene expression of c-Src and EGFR, as well as downstream STAT5 and MAPK-1 with the tumor suppressor gene P53 to all be downregulated. EGFR and c-Src protein expression was also reduced when assessed by western blotting techniques. The effect of the antisense oligonucleotide on HT29 cell proliferation was determined by MTT assay, reduction beijng observed after 48 hours. In summary, nano-drug, anti-EGFR and c-Src specific antisense oligodeoxynucleotides were effectively transferred into HT-29 cells and inhibited gene expression in target cells. Based on the results of this study it appears that the use of antisense EGFR and c-Src simultaneously might have a significant effect on colon cancer growth by down regulation of EGFR and its downstream genes.

• 한국환경농학회지
• /
• 제13권3호
• /
• pp.311-320
• /
• 1994

농축산(農畜産) 폐기물(廢棄物)의 혐기적(嫌氣的) 처리공정(處理工程)에 적용하기 위하여 아한대(亞寒帶) 지역(地域)으로 부터 분리(分離)한 clostridia의 생화학적(生化學的) 특성(特性)을 조사(調査)한 결과(結果)는 다음과 같았다. 1. Cellulose 배지로 저온(低溫)에 집적배양(培養)한 결과 $CH_4$ 생성이 왕성(旺盛) 하였던 늪지 퇴적물(堆積物)(Canada, $56.9^{\circ}N$), 호수퇴적물(堆積物) Ⅳ(Canada, $55.0^{\circ}N$) 그리고 갯벌흙 Ⅱ(Korea, $37.0^{\circ}N$)에서 산생성능(酸生成能)이 우수하고 동시에 메탄을 생성하는 Clostridia, 즉 Clostridium botulinum SRC-64, Clostridium scatologens SRC-91 및 Clostridium tyrobutyricum SRC-100 을 각각 분리(分離)하였다. 분리(分離)된 clostridia 3균주(菌株)의 최적(最適) 생육온도(生育溫度)는 $37^{\circ}C$이었고 최저(最低) 생육온도(生育溫度)는 $10^{\circ}C$ 정도 이었으며, 그리고 이들 균주(菌株)는 특이적 메탄 생성균(生成菌)의 조효소(組酵素)인 $F_{420}$을 함유 하였다. 2. 포도당의 혐기분해(嫌氣分解) 생성물(生成物)로서 SRC-64 는 $H_2$, acetic, isovaleric, caproic acid 를 생성(生成)하였으며, SRC-91 은 $H_2$, propionic, butyric, valeric, caproic acid 를, SRC-100 은 acetic 과 propionic acid 를 주로 생성(生成) 하였다. 또한 3 균주(菌株) 모두 $13^{\circ}C$, 2 일(日) 배양조건(培養條件)에서 2.6-8.68 n moles/ml 범위(範圍)의 $CH_4$ 생성능력(生成能力)을 나타냈다.

• BMB Reports
• /
• 제43권2호
• /
• pp.91-96
• /
• 2010

The function of macrophage inhibitory cytokine-1 (MIC-1) in cancer remains controversial, and its signaling pathways remain poorly understood. In this study, we demonstrate that MIC-1 induces the transactivation of EGFR, ErbB2, and ErbB3 through the activation of c-Src in SK-BR-3 breast cells. MIC-1 induced significant phosphorylation of EGFR at Tyr845, ErbB2 at Tyr877, and ErbB3 at Tyr1289 as well as Akt and p38, Erk1/2, and JNK mitogen-activated protein kinases (MAPKs). Treatment of SK-BR-3 cells with MIC-1 increased the phosphorylation level of Src at Tyr416, and induced invasiveness of those cells. Inhibition of c-Src activity resulted in the complete abolition of MIC-1-induced phosphorylation of the EGFR, ErbB2, and ErbB3, as well as invasiveness and matrix metalloproteinase (MMP)-9 expression in SK-BR-3 cells. Collectively, these results show that MIC-1 may participate in the malignant progression of certain cancer cells through the activation of c-Src, which in turn may transactivate ErbB-family receptors.

• Journal of Welding and Joining
• /
• 제14권3호
• /
• pp.48-57
• /
• 1996

A study was made to examine the characteristics of base metal and stress relief cracking(SRC) of heat affected zone(HAZ) for HY-100 and Cu-bearing HSLA-100 steels. The Gleeble thermal/mechanical simulator was used to simulate the SRC/HAZ. The details of mechanical properties of base plate and SRC tested specimens were studied by impact test, optical microscopy and scanning electron microscopy. The specimens were aged at $650^{\circ}C$ for HSLA-100 steel and at $660^{\circ}C$ for HY-100 steel and thermal cycled from $1350^{\circ}C$ to $25^{\circ}C$ with a cooling time of $\Delta$t_${800^{circ}C/500^{circ}C}$=21sec. corresponds to the heat input of 30kJ/cm. The thermal cycled specimens were stressed to a predetermined level of 248~600MPa and then reheated to the stress relief temperatures of $570~620^{\circ}C$. The time to failure$(t_f)$ at a given stress level was used as a measure of SRC susceptibility. The strength, elongation and impact toughness of base plate were greater in HSLA-100 steel than in HY-100 steel. The time to failure was decreased with increasing temperature and/or stress. HSLA-100 steel was more susceptible to stress relief cracking than HY-100 steel under same conditions. It is thought to be resulted from the precipitation of $\varepsilon$-Cu phase by dynamic self diffusion of solute atoms. By the precipitation of $\varepsilon$-Cu phase, the differential strengthening of grain interior relative to grain boundary may be greater in the Cu-bearing HSLA-100 steel than in HY-100 steel. Therefore, greater strain concentration at grain boundary of HSLA-100 steel results in the increased SRC susceptibility. The activation energies for SRC of HSLA-100 steel are 103.9kcal/mal for 387MPa and 87.6kcal/mol for 437MPa and that of HY-100 steel is 129.2kcal/mol for 437MPa.

• Journal of Ginseng Research
• /
• 제22권2호
• /
• pp.126-132
• /
• 1998

In the present study, we examined effects of ginseng saponins (ginsenosides) on pp60c-src protein tyrosine kinase (PTK) activity, intracellular calcium concentration ([$Ca^{2+}$]i), and cell proliferation in NIH3T3 cells. Eight different ginsenosides [ginsenoside-Rb1 (G-$Rb_1$), -$Rb_2$, -Rc, -Rd, -Re, -Rf, -$Rg_1$, -$Rg_2$) and ginseng total saponin (GTS) were used for these experiments. All ginsenosides and GTS tested stimulated the activation of $pp60^{c-src}$ kinase, and especially G-$Rb_1$,-Rd,-$Rg_1$, and -$Rg_1$ showed a higher stimulatory effect than others at 16.7 $\mu\textrm{g}$/ml of ginsenosides with a 18 hr-incubation, increasing the activity by 4.5, 3.5, 3.5, and 3.0-fold, respectively, over that of untreated control. In addition, both G-Rd and -$Rg_2$)Rg2 increased ($Ca^{2+}$), to 202 and 334 nM, respectively, about 2-3-fold above the basal level within 7min at 250 $\mu\textrm{g}$/yml of ginsenosides. The increases of ($Ca^{2+}$), were eliminated by Pretreatment of EGTA, an extracellular calcium chelator, suggtasting that they result from an influx of calcium ion from extracellular medium rather than an efflux from intracellular calcium store, endoplasmic reticulum (ER). All ginsenosides studied enhanced cell proliferation to 1.2-1.4-fold over that of untreated control at 5~250 $\mu\textrm{g}$/ml of concentrations. Interestingly the promotion of cell proliferation by ginsenosides corresponded with the activation of c-src kinase, which is an early step in the mitogenic signaling cascade. Taken together, we suggest that some ginsenosides may lead to cellProliferation via the activation of cellular signal transduction Pathway involving $pp60^{c-src}$ kinase.

• Journal of Korean Neurosurgical Society
• /
• 제55권5호
• /
• pp.255-260
• /
• 2014

Objective : The purpose of this study was to compare the radiological and neurological outcomes between two atlantoaxial fusion method for atlantoaxial stabilization; C1 lateral mass-C2 pedicle screws (screw-rod constructs, SRC) versus C1-2 transarticular screws (TAS). Methods : Forty-one patients in whom atlantoaxial instability was treated with atlantoaxial fixation by SRC group (27 patients, from March 2005 to May 2011) or TAS group (14 patients, from May 2000 to December 2005) were retrospectively reviewed. Numeric rating scale (NRS) for pain assessment, Oswestry disability index (ODI), and Frankel grade were also checked for neurological outcome. In radiologic outcome assessment, proper screw position and fusion rate were checked. Perioperative parameters such as blood loss during operation, operation time, and radiation exposure time were also reviewed. Results : The improvement of NRS and ODI were not different between both groups significantly. Good to excellent response in Frankel grade is shown similarly in both groups. Proper screw position and fusion rate were also observed similarly between two groups. Total bleeding amount during operation is lesser in SRC group than TAS group, but not significantly (p=0.06). Operation time and X-ray exposure time were shorter in SRC group than in TAS group (all p<0.001). Conclusion : Both TAS and SRC could be selected as safe and effective treatment options for C1-2 instability. But the perioperative result, which is technical demanding and X-ray exposure might be expected better in SRC group compared to TAS group.