• 대한약리학회지
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• 제32권3호
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• pp.417-424
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• 1996

본 연구는 C5a에 의해 자극된 호중구에서 세포내 칼슘유리와 세포외부로부터 칼슘유입에 있어 protein kinase C와 protein tyrosine kinase의 관여 여부를 조사하였다. Protein kinase C 억제제인 staurosporine과 H-7은 C5a에 의해 자극된 호중구에서 세포내 칼슘유리를 억제하였으나, 세포막을 교차한 칼슘유입이나 세포내 칼슘농도 증가에 영향을 나타내지 않았다. C5a에 의한 세포내 칼슘유리와 칼슘유입은 protein tyrosine kinase 억제제인 genistein과 methyl-2,5-dihydroxycinnamate에 의해서 억제 되었다. ADP에 의해 야기된 세포내 칼슘농도의 증가는 genistein과 methyl-2,5-dihydroxycinnamate에 의해서 억제되었으나 staurosporine과 H-7의 영향은 받지 않았다. Genistein과 methyl-2,5-dihydroxy-cinnamate는 thapsigargin을 처리한 호중구에서 칼슘유입을 감소시켰으나 이에 대한 staurosporine 과 H-7의 효과는 나타나지 않았다. 호중구를 phorbol 12-myristate 13-acetate로 전처치하였을때 세포내 칼슘증가에 미치는 C5a의 자극 효과는 감소하였다. 이상의 결과로 부터 protein tyrosine kinase는 C5a에 의해 활성화된 호중구에서 세포내 칼슘유리와 세포막을 교차한 칼슘유입의 조절에 관여할 것으로 추정된다.

• 대한약리학회지
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• 제32권1호
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• pp.103-112
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• 1996

Platelet-activating factor (PAF)에 의하여 자극된 호중구 respiratory burst, 탈과립과 세포질 칼슘농도의 증가에 있어 protein kinase C와 protein tyrosine kinase의 역할을 관찰하였다. PAF에 의하여 자극된 호중구에서 superoxide 및 $H_2O_2$의 생성과 myeloperoxidase와 acid phosphatase의 유리는 protein kinase C 억제제인 staurosporine과 H-7 그리고 protein tyrosine kinase 억제제인 genistein과 tyrphostin에 의하여 억제되었다. PAF에 의한 호중구 세포내 칼슘농도의 증가는 staurosporine, genistein과 methyl-2,5-dihydroxycinnamate에 의하여 억제 되었다. Staurosporine은 PAF에 의하여 자극된 호중구에서 세포내 칼슘유리와 망간유입을 억제 하였다. Genistein과 methyl-2,5-dihydroxycinnamate는 PAF에 의한 망간유입을 억제하였으나, 세포내 칼슘유리에 대한 이들의 효과는 관찰되지 않았다. PMA에 의하여 활성화된 호중구에서 세포내 칼슘농도의 증가에 대한 PAF의 자극효과는 감소되었다. Protein kinase C와 protein tyrosine kinase는 PAF에 의하여 자극된 호중구에서의 respiratory burst, lysosomal enzyme유리와 칼슘동원에 관여할 것으로 제시된다. 세포내 칼슘농도의 증가는 protein kinase의 영향을 다르게 받는 세포내 칼슘유리와 세포외부로 부터의 칼슘유입에 의하여 이루어질 것으로 추정된다. Protein kinase C가 활성화되어 있는 상태에서 세포내 칼슘동원에 대한 PAF의 자극작용은 감소될 것으로 시사된다.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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• pp.216.1-216.1
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• 2003

Neuronal apoptosis may contribute to the pathological neuronal loss in certain disease states such as neurodegenerative diseases. Staurosporine (ST), a nonselective protein kinase inhibitor, has been shown to induce apoptosis in a variety of cells including nerve cell lines. In this study, we investigated the neuroprotective effect of sauchinone, which is a unique lignan from Sauchinone Chinensis, on ST-induced apoptosis in C6 rat glioma cells. (omitted)

• The Korean Journal of Physiology and Pharmacology
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• 제2권3호
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• pp.377-384
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• 1998

Gold compounds depress phagocytic cell responses, including chemotaxis, and respiratory burst. However, the effects of gold compounds on the function of phagocytic cells are variable according to the preparation of medicine. In this study, effect of tetrachloroauric acid on activated neutrophil responses, including respiratory burst, lysosomal enzyme release and change of intracellular $Ca^{2+}$ level and on the synthesis of interleukin-8 and granulocyte-macrophage colony stimulating factor by macrophages was studied. This study further examines how gold compounds affect the activation processes. The respiratory burst stimulated by complement C5a, degraded IgG and PMA in neutrophils was inhibited by tetrachloroauric acid. In contrast to C5a and degraded IgG, PMA-stimulated superoxide production was weakly inhibited by tetrachloroauric acid. Staurosporine, genistein, EGTA and verapamil inhibited superoxide and $H_2O_2$ production caused by C5a and degraded IgG. PMA-stimulated superoxide production was inhibited by staurosporine but was not affected by genistein. Tetrachloroauric acid, genistein, EGTA and verapamil inhibited the release of acid phosphatase and myeloperoxidase, while the effect of staurosporine was not detected. The synthesis of interleukin-8 and granulocyte-macrophage colony stimulating factor by $interleukin-1{\beta}$ in macrophages was inhibited by tetrachloroauric acid. Preincubation with tetrachloroauric acid, genistein, EGTA and verapamil, the elevation of [$Ca^{2+}_i$] evoked by C5a was inhibited. Store-regulated $Ca^{2+}$ entry in thapsigargin-pretreated neutrophils was decreased by the addition of tetrachloroauric acid and genistein. The effect of staurosporine on intracellular $Ca^{2+}$ mobilization was not observed. In conclusion, tetrachloroauric acid may suppress neutrophil responses through its inhibitory action on elevation of intracellular $Ca^{2+}$ level and protein kinase C. It might exhibit an inhibitory effect on the action of protein tyrosine kinase. Tetrachloroauric acid depresses cytokine production by macrophages.

• Asian Pacific Journal of Cancer Prevention
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• 제15권8호
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• pp.3575-3579
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• 2014

TDP-43 is a ubiquitously expressed DNA/RNA binding protein that has recently attracted attention for its involvement in neurodegenerative diseases. While TDP-43 has been found to participate in various important cellular activities including stress and apoptosis, little is known about its role in cancer cells. Here we report that staurosporine (STS) induced apoptosis in U87 glioma cells is associated with rapid downregulation of TDP-43 at both mRNA and protein levels. The latter is dependent on activation of caspase 3. More importantly, we have shown that knockdown of TDP-43 by specific siRNA dramatically enhanced cytotoxicity of STS. These results suggest that normal level of TDP-43 may be protective for cancer cells under apoptotic insult.

• 한국동물학회지
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• 제38권2호
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• pp.286-293
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• 1995

The present studies were undertaken to examine the activitites of PKC isoforms in cultures of chick limb mesenchvme. Micromass cultures were prepared using wing buds of stage 23/24 (Hamburger and Hamilton, 19511 chick embryo. The cells were homogenized and DEAE-cellulose column chromatography was performed to get fraction containing protein kinase C (PKC) activity. PKC isoforms were resolved with hvdroxyapatitie column chromatography. Profile of PKC isoforms of cultures were compared with that of rat brain. Activity of $PKC-\beta$ isoform was appeared at the early stage of chondrogenesis. On 3 daw of culture, activities of both PKC a and $\beta$ were observed with remarkable increase but no activity of y isoform was appeared. Treatment of phorbol-12-mvristate-13-acetate (PMA) (10-7 M) to the culture inhibited chondrosenesis and down-regulated a and $\beta$ isoforms. Staurosporine promoted chondro!genesis without any effect on PKC isioforms profile. These data indicate that PKC a and $\beta,$ especiallv $\beta$ isoform is related to chondrosenesis and the promoting effect of staurosporine on chondrogenesis is not related to PKC isoforms activities.

• Animal cells and systems
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• 제12권4호
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• pp.211-217
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• 2008

Bax, a pro-apoptotic member of Bcl-2 family proteins, plays a central role in the mitochondria-dependent apoptosis. Apoptotic signals induce the translocation of Bax from cytosol into the mitochondria, which triggers the release of apoptogenic molecules such as cytochrome C and apoptosis-inducing factor, AIF. Bax-inhibiting peptide(BIP) is a cell permeable peptide comprised of five amino acids designed from the Bax-interaction domain of Ku70. Because BIP inhibits Bax translocation and Bax-mediated release of cytochrome C, BIP suppresses Bax-dependent apoptosis. In this study, we observed that BIP inhibited staurosporine-induced neuronal death in cultured cerebral cortex and cerebellar granule cells, but BIP failed to rescue granule cells from trophic signal deprivation-induced neuronal death, although both staurosporine-induced and trophic signal deprivation-induced neuronal death are dependent on Bax. These findings suggest that the mechanisms of the Bax activation may differ depending on the type of cell death induction, and thus BIP exhibits selective suppression of a subtype of Bax-dependent neuronal death.

• Archives of Pharmacal Research
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• 제22권2호
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• pp.119-123
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• 1999

Parathyroid hormone (PTH) treatment of bone and kidney-derived cells not only activates adenyly cyclase buy also increases intracellular free calcium, and translocates protein kinase C (PKC) from cytosol to plasma membranes. We have found that acute phorbol ester pretreatment significantly decreases PTH-induced calcium transients and the effect of phorbol ester was antagonized by staurosporine (ST). Although the major effect of ST in that study was the reversal of the action of phorbol ester, it appeared that ST may also have promoted the effect of PTH directly. To further investigate the observation, we examined the effect of ST on the intracellular calcium transients induced by PTH and $\alpha$-thrombin ($\alpha$-TH). For calcium transient experiments, UMR-106 cells were loaded with 2 mM fluo-acetoxymethylester for 30 min at room temperature. The cells were then washed and suspended in buffer containing 1 mM calcium. Fluorescence was detected at 530 nm, with excitation at 505 nm. ST alone did not cause calcium transients, but enhanced the transients elicited by PTH response. added 5 min before the hormone. Another protein kinase inhibitor H-7 likewise enhanced the calcium responses elicited by PTH, while genistein did not affect PTH response. Calcium transients elicited by $\alpha$-TH were also enhanced by ST. The results suggest that there might be tonically activated endogenous protein kinase(s) which inhibit calcium signaling of some calcemic agents.

• 한국식품영양과학회지
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• 제46권2호
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• pp.161-168
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• 2017

본 연구에서는 산화적 스트레스로 유도한 시신경 세포사멸에 대한 차조기 물 추출물(PFE)의 효과를 확인하였다. Staurosporine으로 분화된 ssdRGC-5 세포에 buthionine과 glutamate(B/G)로 산화적 스트레스를 유도하였으며, LDH release assay, MTT reduction assay를 통하여 PFE가 농도 의존적으로 B/G에 의한 세포사멸을 억제함을 관찰하였다. 세포사멸의 기전을 연구하기 위해 caspase 활성, 세포 내 ROS 생성량, 세포고사 관련 단백질 발현을 관찰한 결과, B/G에 의해 증가한 ROS 생성량, caspase 활성을 PFE가 억제하였고, 세포질로 방출된 cytochrome c와 미토콘드리아로 이동한 Bax도 감소함을 확인하였다. 이상의 결과로부터 차조기는 산화적 스트레스로 유도된 시신경 세포사멸 과정에서 항산화 효과와 미토콘드리아성 세포사멸을 완화함으로써 세포 보호 작용을 나타냄을 확인하였다.

• 고려인삼학회:학술대회논문집
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• 고려인삼학회 1998년도 Advances in Ginseng Research - Proceedings of the 7th International Symposium on Ginseng -
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• pp.231-243
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• 1998

In the present study, we provide evidence that ginsenoside $Rh_2$ (G-$Rh_2$) as well as staurosporine induces apoptosis of human hepatoma SK-HEP-1 cells by caspase 3-mediated processing of $p21^{WAFI/CIPI}$ in the early stage of apoptosls. Immunoblottings showed that G-$Rh_2$ as well as statrosporine induced the processing of caspase-3 to an active form, pl7. In stable Bcl-2 transfectants however, G-$Rh_2$ induced DNA fragmentation, while staurosporine did not. In the early stage of apoptosis, $p21^{WAFI/CIPI}$ was detected to undergo proteolytic processing specifically conducted by caspase-3. $p21^{WAFI/CIPI}$ translated in vitro was cleaved into a p14 fragment, when incubated with cell extracts obtained from either G-$Rh_2$- or staurosporine-treated cells. Cleavage was equally inhibited in both cases by adding Ac-DEVD-cho, a specific caspase-3 inhibitor, but not by Ac-YVkD-cho, a specific caspase-l inhibitor. Similarly, $p21^{WAFI/CIPI}$ was efficiently leaved by recombinant caspase-3 overexpressed in E. coli. Moreover, the endogenous $p21^{WAFI/CIPI}$ of untreated-cell extracts was also cleaved by recombinant caspase-3. Mutation analysis allowed identification of two caspase-3 cleavage sites, $DHVD^{112}$/L and $SMTD^{149}$/F, which are located within, or near the interaction domains for cyclins, Cdks, and PCNA. Taken together, these results show that G-$Rh_2$ as well as staurosporine increases caspase-3 activity, which in turn directly cleaves $p21^{WAFI/CIPI}$ resulting in elevation of Cdk kinase activity in the early stages of apoptosis. We propose that proteolytic cleavage of $p21^{WAFI/CIPI}$ is a functionally relevant event that allows unleashing the cyclin/Cdk activity from the inhibitor seen in the earlier stage of apoptosis, the event of which may be associated with the triggering mechanism for the execution of apoptosis.