• Asian Pacific Journal of Cancer Prevention
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• 제15권17호
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• pp.7245-7249
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• 2014

Glucose regulated protein 78 (GRP78) is usually recognized as a chaperone in the endoplasmic reticulum. However, increasing evidence indicates that GRP78 can be translocated to the cell surface, acting as a signaling receptor for a variety of ligands. Since little is known about the secretion of GRP78 and its role in the progression of colon cancer we here focused on GRP78 from colon cancer cells, and purified GRP78 protein mimicking the secreted GRP78 was able to utilize cell surface GRP78 as its receptor, activating downstream PI3K/Akt and Wnt/${\beta}$-catenin signaling and promote colon cancer cell proliferation. Our study revealed a new mode of action of autocrine GRP78 in cancer progression: secreted GRP78 binds to cell surface GRP78 as its receptor and activates intracellular proliferation signaling.

• BMB Reports
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• 제47권8호
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• pp.445-450
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• 2014

Glucose regulated protein 78 (GRP78) is frequently highly expressed in tumor cells, contributing to the acquisition of several phenotypic cancer hallmarks. GRP78 expression is also positively correlated with tumor metastasis, and promotes hepatocellular carcinoma cell invasion via increasing cell motility, however, other mechanisms involving the prometastatic roles of GRP78 remain to be elucidated. Here we report that forced GRP78 expression promotes colon cancer cell migration and invasion through upregulating MMP-2, MMP-9 and especially uPA production. These effects of GRP78 are mediated by enhancing the activation of ${\beta}$-catenin signaling. Interestingly, we identify that GRP78 interacts with uPA both in the cells and in the culture medium, suggesting that GRP78 protein is likely to directly facilitate uPA secretion via protein-protein interaction. Taken together, our findings demonstrate for the first time that besides stimulation of cell motility, GRP78 can act by increasing proteases production to promote tumor cell invasion.

• 한국동물학회지
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• 제38권2호
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• pp.167-176
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• 1995

Glucose-regulated proteins (grp's), srp94 3nd grp78/BiP, are a group of stress proteins which are highly synthesized in cells exposed to a variety of stressful agents including tunicamycin 3nd Ca2+ ionophore. Grp78/BiP is hon to function as a molecular chaperone which regulates the folding and assembly of secretory or membrane proteins, but the biological function of grp941 remains to be elucidated. In this study, we have examined the intracellular distribution of grV's and the function of srp94. Grp's are resident in the endoplasmic reticulum (ERI 3nd a specific sequence (Lys-Asp-Glu-Leu) at their C-terminus is known to be responsible for their retention within the ER. However, it has been unclear whether upon disturbance of cellular Caa+ homeostasis by the Ca2+ ionophore, grp94 is retained within the ER or secreted into the medium. In this study, we showed that in the presence of C3a+ ionophore, grp94 and gif78/BiP are present in the cells, mainly within the ER. We have also investigated whether grp94 might function as a molecular chaperone. Here we showed that in the immunoglobulin (Ig)-secreting hvbridom3 cells, grp94 transientlY interacts with fully glycosylated Is heavy chain, suggesting that grpg94 may be involved in facilitating the folding and assembly of Ig heavy chains.

• 한국동물학회지
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• 제37권2호
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• pp.137-143
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• 1994

Microiniection of genes Into Xenopus laeuis oocvtes in highly useful in the annvsis of gene regulation, since a large number of oocvtes can be injected in a relatively short time. The GRP78 enhancer has been identified to a 291-bp fragment that spans a region of GRP78 promoter between -378 and -87 (Lin et at., 1986: Kim and Lee, 1989). We examined whether this GRP78 enhancer is effective in directing expression of heterologous gene in Xenopus laeuis oocytes. The chloramphenicol acetvltransferase (CAT) fusion constructs containing the GRP78 promoter and the SV4O early promoter were constructed and were injected into nuclei of Xencpus laeuis oocvtes. The recipient oocvtes were then assayed for CAT activity. The fusion constructs exhibited higher activity as compared to SV40 promoter tested here. The GRP78 enhancer showed 8.5- to 9.2-fold enhancement over that of the SV4O promoter. The orientation of GRP78 enhancer with respect to the direction of CAT transcription unit had no significant effect. Thus, the GRP78 enhancer is a viable candidate for the construction of expression system for use in Xenopus laevss oocvtes and will be important for the studY of a gene expression throughout development.

• Asian Pacific Journal of Cancer Prevention
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• 제16권8호
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• pp.3517-3522
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• 2015

Background: Glucose regulated protein 78 (GRP78) is a type of molecular chaperone. It is a possible candidate protein that contributes to development of drug resistance. We first examined the involvement of GRP78 in chemotherapy-resistance in human ovarian cancer cell. Materials and Methods: The expression of GRP78 mRNA and protein were examined by RT-PCR and western blotting, respectively, in human ovarian cancer cells line (HO-8910). Sensitivity of HO-8910 to paclitaxel was determined with methyl thiazolyl tetrazolium (MTT). Suppression of GRP78 expression was performed using specific small-interfering RNA (siRNA) in HO-8910 cells, and cell apoptosis was assessed by flow cytometry. Statistical analysis was performed using the SPSS 15.0 statistical package. Results: HO-8910 cells, with high basal levels of GRP78, exhibited low sensitivity to paclitaxel. The mRNA and protein levels of GRP78 were dramatically decreased at 24h, 48h and 72h after transfection and the sensitivity to paclitaxel was increased when the GRP78 gene was disturbed by specific siRNA transfection. Conclusions: The results suggested that high GRP78 expression might be one of the molecular mechanisms causing resistance to paclitaxel, and therefore siRNA of GRP78 may be useful in tumor-specific gene therapy for ovarian cancer.

• 대한의생명과학회지
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• 제11권3호
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• pp.311-318
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• 2005

Grp78, discovered as one of the putative target genes of Hoxc8, is a highly conserved stress protein and functions as a molecular chaperone in the endoplasmic reticulum (ER). In order to see the stage-specific expression pattern of Grp78 during development, mouse embryos from day 7.5 to 17.5 p.c. were isolated, and RT-PCR as well as in situ hybridization was performed. When RT-PCR was performed using Grp78 specific primers, periodic expression pattern was detected. And also a region-specific expression pattern was detected with a strong expression in the trunk part of day 11.5 p.c. embryo, like that of Hoxc8. When in situ hybridization was performed, Grp78 was revealed to be expressed in the endoderm, somite, neuroepithelium cells of neural tube in early embryos. In the case of late embryos, Grp78 expression was detected in the liver, segmental bronchus within cranial lobe of lung, ossification center within the cartilage primordium of rib and vertebra, submandibular gland, as well as metanephros. These expression patterns are very much similar to those of Hoxc8. Since Hoxc8 has been reported to regulate apoptosis during organogenesis, it might be possible that the apoptotic function could have been conveyed through the expression of Grp78, implying that the Grp78 is one of the Hoxc8 downstream target genes.

• 생명과학회지
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• 제15권1호
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• pp.87-93
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• 2005

본 연구에서는 Streptozotocin-유발 당뇨 쥐를 대상으로 8주 동안 운동 강도에 따른 혈당, 인슐린, 골격근 내 GLUT-4 발현 량과 GRP-78 발현량 변화를 관찰함으로써 당뇨 쥐의 혈당을 낮추는데 효과적인 운동 강도는 무엇이며, 기전적인 원인은 무엇인지를 밝히고자 실시되었다. 본 연구에서 Streptozotocin으로 당뇨를 유발시킨 흰쥐를 대상으로 운동 강도 처치에 따른 GLUT-4발현 량을 비교한 결과, 기본적으로 당뇨가 유발된 흰쥐의 GLUT-4 발현 량은 정상 쥐에 비해 현저하게 감소하는 것을 볼 수 있었다. 이중 8주 동안 저강도 운동 수행 한 쥐들의 경우는 정상 쥐 수준에 가까운 발현 량을 관찰 할 수 있었으며, 흰쥐 가자미 근 내 GRP-78 발현 량은 당뇨 유발 쥐의 경우 정상 쥐의 유의하게 증가한 것을 관찰할 수 있었고, 저강도 운동을 수행한 집단에서의 GRP-78 발현 량은 정상 흰쥐에서 발현되는 양에 비해 유의하게 높은 수준을 보이기는 했지만 당뇨 유발 대조군과 고강도 운동 군에 비해서는 유의하게 낮은 수준으로 나타났다. 결론적으로 저강도의 운동은 당뇨 질환자의 혈당을 낮추는 데 가장 효과적인 운동이라 할 수 있으며, 이러한 결과의 기전적인 원인은 저강도의 운동이 근육 내 GLUT-4의 증가로 세포내 당 유입을 촉진시킴으로 해서 나타난 사실이라 생각되며 이 같은 사실은 GRP-78의 발현 량의 감소로 확인할 수 있었다.

• 대한의생명과학회지
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• 제17권1호
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• pp.1-5
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• 2011

Previously, we have identified 14 putative downstream target genes of Hoxc8 homeoprotein in F9 murine embryonic teratocarcinoma cells through proteomics analysis. Among those, we tested a possibility of a DNA-k type molecular chaperone, Grp78, as a direct downstream target of Hoxc8, by cloning a 2.4 kb upstream region of murine Grp78 into a reporter plasmid and by testing if Hoxc8 can regulate its expression. We observed that Hoxc8 proteins could transactivate the reporter gene, which was affected by small interference RNAs (siRNAs) against to Hoxc8, suggesting that Grp78 is a novel downstream target of Hoxc8 in vivo.

• 한국발생생물학회지:발생과생식
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• 제21권4호
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• pp.407-415
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• 2017

In the present study, we investigated the role of binding immunoglobulin protein/glucose-regulated protein, 78-kDa (BIP/GRP78)-regulated endoplasmic reticulum (ER)-stress on meiotic maturation and cumulus cells expansion in porcine cumulus-oocyte complexes (COCs). Previously, it has been demonstrated that unfolded protein response (UPR)-related genes, such as molecules involved in ER-stress defense mechanisms, were expressed in matured oocytes and cumulus cells during in vitro maturation (IVM) of porcine oocytes. However, BIP/GRP78-mediated regulation of ER stress in porcine oocytes has not been reported. Firstly, we observed the effects of knockdown of BIP/GRP78 (an UPR initiation marker) using porcine-specific siRNAs (#909, #693, and #1570) on oocyte maturation. Among all siRNAs, siRNA #693 significantly reduced the protein levels of UPR marker proteins (BIP/GRP78, ATF4, and P90ATF6) in porcine COCs observed by Western blotting and immunofluorescence analysis. We also observed that the reduction of BIP/GRP78 levels by siRNA#693 significantly inhibited the meiotic maturation of oocytes (siRNA #693: $32.5{\pm}10.1%$ vs control: $77.8{\pm}5.3%$). In addition, we also checked the effect of ER-stress inhibitors, tauroursodeoxycholic acid (TUDCA, $200{\mu}M$) and melatonin ($0.1{\mu}M$), in BIP/GRP78-knockdown oocytes. TUDCA and melatonin treatment could restore the expression levels of ER-stress marker proteins (BIP/GRP78, $p-eIF2{\alpha}$, $eIF2{\alpha}$, ATF4, and P90ATF6) in siRNA #693-transfected matured COCs. In conclusion, these results demonstrated that BIP/GRP78-mediated regulation of UPR signaling and ER stress plays an important role in in vitro maturation of porcine oocytes.

• 생명과학회지
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• 제14권5호
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• pp.741-746
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• 2004

본 연구는 운동강도 차이에 따른 카페인 구강 투여가 STZ-유발 당뇨 쥐 가자미근에서 GLUT-4와 GRP-78 단백질 발현에 미치는 영향을 규명하기 위하여 F344계 수컷 횐쥐를 무작위 표본추출에 의하여 당뇨유발군(n=6), 당뇨유발-카페인 투여군(n=6), 당뇨유발-카페인투여 저강도운동군(n=6), 당뇨유발-카페인투여 중강도운동군(n=6), 그리고 당뇨유발-카페인투여 고강도 운동군(n=6)으로 분류하였다. 저강도 운동은 트레드밀 경사도 0%에서 8 m/min 속도로, 중강도 운동은 트레드밀 경사도 0%에서 16 m/min 속도로, 고강도운동은 트레드밀 경사도 0%에서 25 m/min속도로 30분간 1회 운동을 실시하였다. GLUT4단백질 발현은 당뇨군에 비해서 당뇨유발군-카페인 투여군과 당뇨유발-카페인투여 저강도 운동군에서 차이가 없었으며, 당뇨유발-카페인투석 중강도 운동군에서는 다소 감소하였으나 당뇨유발-카페인투여 고강도 운동군에서 증가하였다. GRP-78 단백질 발현은 당뇨군에 비해서 당뇨유발-카페인투여 저강도 운동군, 당뇨유발-카페인투여 중강도 운동군, 그리고 당뇨유발-카페인투석 고강도 운동군에서 감소하였으나, 당뇨유발-카페인 투여군에서는 다소 증가한 것으로 나타났다 고강도 일회성 운동이 인슐린 민감도를 개선시켜 인슐린 요구량을 낮추는데 이러한 효과는 내형질세망에서 세포막으로의 GLUT-4 단백질의 전이와 GLUT-4 단백질 양의 증가 때문이다. 운동군에서의 GRP-78 단백질이 감소된 기전은 정확히 밝힐 수는 없지만, 카페인으로 인한 지질 동원이 운동 시 작업근의 세포에 많은 에너지를 공급하여 세포가 받는 스트레스를 완화시켜 주었기 때문이라고 추측된다.