• BMB Reports
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• 제29권1호
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• pp.73-80
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• 1996

The effect of albumin on phosphatidylcholine (PC) metabolism in Hep-G2, 3T3-H.ras, and 3T3 cells pre-labelled with [Me-$^3H$]choline was studied. The [$^3H$]choline was more efficiently taken up and incorporated into cellular phospholipids in 3T3-H.ras cells than in Hep-G2 and 3T3 cells. In each of the three cell lines, most of the [$^3H$]choline metabolized into the phospholipids was incorporated into PC and only minor was incorporated into lysophosphatidylcholine (LPC). Bovine serum albumin stimulated the release of [$^3H$]LPC and [$^3H$]PC from each of the three cell lines pre-labelled with [$^3H$]choline. [$^3H$]PC was also released in the absence of albumin but [$^3H$]LPC was not. The efficiency of LPC secretion represented as the proportion of medium [$^3H$]LPC to cellular [$^3H$]choline lipid during a chase period is approximately 9 to 14 times greater in 3T3 cells compared with the transformed 3T3-H.ras and Hep-G2 cells. A similar comparison of published data for rat hepatocytes with Hep-G2 shows secretion to be 35~75 times greater from the rat hepatocytes than from Hep-G2. Also, PC secretion from 3T3 cells was 1.6 times more effective than from 3T3-H.ras, whereas rat hepatocytes secrete PC 2.8~3.8 times more effectively than does Hep-G2. The measurement of specific radioactivity of cellular PC in pre-labelled 3T3 cells showed it to be similar to that of the secreted PC. However, the specific radioactivity of secreted LPC was markedly lower than that of the cellular PC, which suggests that LPC is being secreted from a PC pool distinct from that used for PC secretion.

• 한국식품저장유통학회지
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• 제22권1호
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• pp.19-26
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• 2015

본 연구는 3,3'-diindolylmethane(DIM)이 인간전립선암 세포인 PC3 세포와 DU145 세포의 부착, 이동, 침윤에 미치는 영향을 살펴보았다. DIM은 PC3와 DU145 세포의 부착을 농도 의존적으로 억제하였다. 24시간동안 DIM으로 PC3 세포를 전 처리한 후 부착실험을 한 결과 농도 의존적으로 억제하였다. 그러나 암세포가 부착 시 DIM의 암세포 부착 억제가 전처리에 의한 부착 억제보다 효과적이었다. DIM은 인간 전립선암세포의 이동과 침윤도 억제하였으며 24시간 동안 PC3 세포를 DIM으로 전처리를 했을 때도 침윤 억제효과를 나타내었다. 또한 DIM의 억제효과는 세포주에 따라 다소 다른 경향을 보여 PC3 세포에 대한 억제효과가 DU145 세포에 대한 억제효과보다 큰 것으로 관찰되었다. DIM은 $150{\mu}M$ 까지 세포 독성이 관찰되지 않은 것으로 보고되고 있으므로 본 연구결과 DIM은 세포 독성이 없는 수준에서 인간 전립선암 세포인 PC3와 DU145 세포의 부착, 이동, 침윤을 효과적으로 억제하여 전립선암 전이 억제제로 이용될 수 있을 것으로 사료된다.

• 대한수의학회지
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• 제46권2호
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• pp.87-96
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• 2006

We established an in vitro experimental system using the following procedure. We first introduced tritium-labeled norepinephrine ([$^3$H]-NE) into PC12 cells, The [$^3$H]-NE incorporated into PC12 cells were then stimulated by a high concentration (60 mM) of $K^+$ buffer during 12 minutes. Then, we collected $100{\mu}l$ supernatant and counted the amount of [$^3$H]-NE release from PC12 cells with a scintillation counter. After screening fungal, Streptomyces spp. or bacterial product using this experimental sytem, we obtained FS11052 from Streptomyces spp. which inhibited [$^3$H]-NE release from PC12 cells. FS11052 also inhibits the release of ATP as a neurotransmitter of PC12 cells and rat cortical neurons, The inhibitory effect was seen even when the PC12 cells were treated with low $K^-$ buffer containing ionomycin ($1{\mu}M$) as an ionopore. This result suggests that the inhibitory action of FS11052 on neurotransmitter release appeared after the influx of $Ca^{2+}$.

• Toxicological Research
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• 제14권3호
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• pp.341-348
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• 1998

We established an in vitro experimental system using the following procedure. We first introduced tritium-labelled norepinephrine ([3H]-NE)into PC12 cells. The [3H]-NE incorporated into PC12 cells were then stimulated by a high concentration (60 mM) of $K^+$ during 12 minutes. Then, we counted the amount of [3H]-NE release from PC12 cells with the scintillation counter. After screening fungal, Streptomyces or bacterial product using this experimental system, we obtained S9940 from Streptomyces spp. which inhibited [3H]-NE release from PC12 cells. S9940 also inhibits the release of ATP as a neurotransmitter of PC12 cells and rat cortical neurons. The inhibitory effect was seen even when the PC12 cells were treated with low $K^+$ buffer containing ionomycin $(1\muM)$ as an ionopore. This result suggests that the inhibitory action of S9940 on neurotransmitter release appeared after the influx of $Ca^{2+}$.

• Journal of Nutrition and Health
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• 제41권3호
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• pp.224-231
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• 2008

본 연구는 십자화과 채소의 섭취로 체내유용 물질인 인돌이 전립선암 세포 PC3 cell의 항전이 효과 기전에 미치는 영향에 대하여 알아보았다. 인돌은 전립선암 세포중식을 농도 의존적으로 억제하였으며 인돌에 의한 세포 사멸의 영향과 관계없이 MMP-2, -9의 활성과 전사수준 및 단백질 발현을 억제하였다. 역으로 MMP활성 억제 물질인 TIMP-1,-2의 발현이 인돌 첨가에 의해 증가하였다. $NF{-\kappa}B$의 upstream에 존재하는 MAPK signaling 유전자인 ERK1/2, p38, JNK 발현이 인돌처리로 인산화를 억제하였다. 그리고 전립선암 세포 PC3 침윤성이 인돌 처리 시 유의적으로 감소하였다. 결론적으로 인돌은 PC3 인체 전립선암 세포의 전이 과정을 MAPK phthway를 통한 MMP 활성과 발현 억제, TIMP 발현 증가로 암 세포 전이 억제를 하는 것 으로 나타나 암 전이 억제 식품으로 가능성을 제시한다.

• Journal of Microbiology and Biotechnology
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• 제19권5호
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• pp.530-536
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• 2009

Cobalt chloride ($CoCl_2$) treatment of cells in vitro has been shown to induce cellular changes that are similar to those seen following hypoxia. To identify genes that are differentially expressed in response to treatment with $CoCl_2$, we compared the mRNA expression profiles of PC-3 cells that were treated with $CoCl_2$ with those of untreated PC-3 cells, using specific arbitrary primers and two anchored oligo(dT) primers provided in the ACP-based GeneFishing kits. The results of this study demonstrated that the puromycin-sensitive aminopeptidase (PSA) gene was down regulated in PC-3 cells that were treated with $CoCl_2$. This downregulation of PSA expression, in turn, suppressed the proliferation, migration, and invasion of PC-3 cells, as well as the secretion and expression of matrix metalloproteinase-9 (MMP-9).

• 생물정신의학
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• 제5권2호
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• pp.219-226
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• 1998

Cytosine arabinoside(AraC) inhibits DNA synthesis and ${\beta}$-DNA polymerase, an enzyme involved in DNA repair. This, a potent antimitotic agent, is clinically used as an anticancer drug with side effect of severe neurotoxicity. Earlier reports suggested that inhibition of neuronal survival by AraC in sympathetic neuron may be due to the inhibition of a 2'-deoxycytidine-dependent process that is independent of DNA synthesis or repair and AraC induced a signal that is triggers a cascade of new mRNA and protein synthesis, leading to apoptotic cell death in cultured cerebellar granule cells. The present study would suggest whether caspase family(ICE/CED-3-like protease) involved in AraC-induced apoptosis pathway of PC12 cells. It was observed that treatment of PC12 cells with AraC led to decrease of viability by MTT assay and morphology changes, which did not suggest that AraC induced apoptosis in PC12 cells. The mRNA of caspase-1/caspase-3 were expressed in PC12 cells constitutively, and AraC did not activate caspase family. These results suggest that caspase-1/caspase-3 may not be required for AraC-induced cell death pathway in PC12 cells.

• 대한한의학회지
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• 제24권2호
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• pp.179-192
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• 2003

Objectives : Boyanghwanoh-tang (Buyanhaiwu-tang) has been used as a prescription for stroke, senile and vascular dementia, ischemic brain and heart damage in Oriental traditional medicine. However, there is little known about the mechanism by which the water extracts of Boyanghwanoh-tang (Buyanhaiwu-tang) rescue cells fromthese damages, and little is known about the protective mechanisms of Boyanghwanoh-tang (Buyanhaiwu-tang) on oxidative stress in neuronal cells. Therefore, we have investigated the role of Boyanghwanoh-tang (Buyanhaiwu-tang) on serum and glucose deprived apoptosis in PC12 cells. Methods : PC12 Cells have been used extensively as a model for studying the cellular and molecular effects of neuronal cells. The viability of cells was measured by MIT assay. We used DNA fragmentation and caspase 1, 2, 3, 6, 9-likeproteases activation assay. Transcriptional activation of NF-kB was assessed by using electrophoretic mobility shift assay. Results : Boyanghwanoh-tang (Buyanhaiwu-tang) rescued PC12 cells from apoptotic death by serum and glucose deprivation in a dose-dependent manner. The nuclear staining of PC12 cells clearly showed that Boyanghwanoh-tang (Buyanhaiwu-tang) attenuated nuclear condensation and fragmentation, which represent typical neuronal apoptotic characteristics. Boyanghwanoh-tang (Buyanhaiwu-tang) also prevents fragmentation of genomic DNA and activation of caspase 3-like protease in serum and glucose deprived PC12 cells. Furthermore, Boyanghwanoh-tang (Buyanhaiwu-tang) reduced the activation of NF-kB by serum and glucose-deprived apoptosis. Conclusions : These findings suggest that serum and glucose deprivation induces reduced glutathione (GSH) depletion, and consequently, apoptosis through endogenously produced reactive oxygen species in PC12 cells. Also, our data indicated that Boyanghwanoh-tang (Buyanhaiwu-tang) has protective effects against the serum and glucose deprived deaths of PC12 cells, which are mediated by the generation of GSH that, in turn, can reduce oxidative stress caused by reactive oxygen species (ROS) such as hydrogen peroxide.

• Asian Pacific Journal of Cancer Prevention
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• 제15권23호
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• pp.10079-10083
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• 2015

Background: Endothelin-1 and Endostar are both significant for the progression, proliferation, metastasis and invasion of cancer. In this paper, we studied the effect of ET-1 RNAi and Endostar in PC-3 prostatic cancer cells. Materials and Methods: The lentiviral vector was used in the establishment of ET-1 knockdown PC-3 cells. Progression and apoptosis were assessed by CKK-8 and flow cytometry, respectively. Transwell assay was used to estimate invasion and signaling pathways were studied by Western blotting. Results: ET-1 mRNA and protein in ET-1 knockdown PC-3 cells were reduced to 26.4% and 22.4% compared with control group, respectively. ET-1 RNAi and Endostar both were effective for the suppression of progression and invasion of PC-3 cells. From Western blotting results, the effects of ET-1 regulation and Endostar on PC-3 cells were at least related to some signaling pathways involving PI3K/Akt/Caspase-3, Erk1/2/Bcl-2/Caspase-3 and MMPs (MMP-2 and MMP-9). Furthermore, combined treatment of ET-1RNAi and Endostar was found to be more effective than single treatment. Conclusions: Both ET-1 RNAi and Endostar can inhibit the progression and invasion of PC-3 cells, but combined treatment might be a better therapeutic schedule.

• 생명과학회지
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• 제20권7호
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• pp.991-998
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• 2010

저산소상태(hypoxia)는 세포유형 및 성장조건에 따라 apoptosis를 유발하거나 또는 apoptosis의 진행을 억제한다. 저산소상태 유발물질인 cobalt chloride ($CoCl_2$) 역시 여러 세포 유형에서 apoptosis를 유발한다고 알려져 있으나 그 기작은 불명확하다. 본 연구에서는 PC-3 세포에서 $CoCl_2$에 의해 down-regulation되는 puromycin-sensitive aminopeptidase (PSA)의 세포 내 기능을 조사하였다. 본 연구 결과, PC-3 세포에 puromycin을 처리한 결과 전체 세포 집단의 약 42%에서 apoptosis가 유도되었다. PSA가 apoptosis에 관여하는지를 확인하고자 PSA siRNA로 내재성 PSA의 발현을 억제시킨 다음 apoptosis 연관 특성을 조사한 결과 PSA의 발현 억제에 의해 효과적으로 apoptosis가 유도되었다. 이러한 결과는 PC-3 세포에서 PSA가 세포생존에 중요한 역할을 할 것임을 보여주는 것이라 사료된다.