• Title/Summary/Keyword: P388 cell

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Studies on the Induction of Transformation and Multiplication in Orchid Plants.(III) Expression of Gene Transferred into Orchid Protoplasts by Electroporation (난과식물의 형질전환 유도 및 다량증식에 관한 연구. III. Electroporation에 의해서 자란의 원형질체로 도입된 유전자의 발현)

  • 이정석;황성진김영준황백
    • KSBB Journal
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    • v.6 no.4
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    • pp.385-388
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    • 1991
  • Embryogenic cell suspension cultures of B. striata were established as transferred selected embryogenic callus into liquid medium. Protoplasts isolated from embryogenic cell suspensions were electroporated in buffered solutions containing plasmid DNA of pBI121. Transient GUS (beta-glucuronidase) activity measurement and selection for kanamycin resistent showed that expression of foreign genes and stable transformation were achieved. GUS transient gene expression was increased by increasing DNA concentration of pBI121 plasmid and affected by the level of the applied voltage. An optimal level of GUS activity was obtained after electroporation with a pulse of 200-300 voltage/1180 uF. Protoplast viability was up to the 80% at the optimal voltage.

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Synthesis and Antitumor Activity of Novel Gericudranin E Derivatives (새로운 항암성 제리쿠드라닌 E 유도체의 합성 및 항암활성)

  • 박재호;박경란;호현순;김희두;표명윤
    • YAKHAK HOEJI
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    • v.43 no.5
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    • pp.559-565
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    • 1999
  • The two gericudranin E derivatives, GER-I & II, were synthesized and evaluated their antitumour activities for the elucidation of structure-activity relationship. 2,4,6-Trihydroxyacetophenone was converted to target molecules GER-I and GER-B in 5 steps via sequential protection, aldol condensation, Michael type-cyclization, regioselective C-benzylation. The cellular growth inhibition of compounds GER-I and GER-II were investigated against P388, L1210, K562, HCT-15, SK-HepG-1, MCF-7 as cancer cell lines and mouse splenocytes as a normal cell by MTT assay.

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Purification and Characterization of Mitogen -Activated Protein (MAP) Kinase from Mammalian Tissue Cells (동물 조직세포로부터 Mitogen-activated Protein (MAP) Kinase의 분리 및 성격규명)

  • 김태우;정동주;김윤석
    • Biomedical Science Letters
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    • v.2 no.1
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    • pp.21-30
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    • 1996
  • MAP kinases are a family of serine/threonine specific protein kinases becoming activated in response to different proliferative stimuli by phosphorylation at both threonine and tyrosine residue. Present study shows that MAP kinase was purified from P388 murine leukema cells by SP sephadex C-50, phenyl superose and Mono Q column chromatography and identified with anti-ERKl antibody by western blotting. Immnublotting analysis to the crude extract of P388 cell lysate shows 44 kD and other minor bands but partial purified fraction eluted from phenyl supherose column have 44kD and 66 kD isoform. Subcloned GST-fusion protein from N-terminal of $p56^{kk}$ was tested as a substrate for MAP kinase phosphorylation. It was showed that the wild type and mutant forms(S42A) were fully phosporylated by purified MAP kinase fraction as com-pare with the other mutant form(S59A). This finding suggest that those GST-fusion proteins may be used as substrate for the in vitro test of MAP kinase.

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Studies on the Biological Behaviors of Taxol Derivatives (Taxol 유도체들의 생물학적 거동에 관한 연구)

  • Awh, Ok-Doo;Yoo, Dae-Wung;Im, Sang-Moo
    • The Korean Journal of Nuclear Medicine
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    • v.31 no.4
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    • pp.440-451
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    • 1997
  • This study was designed to prospect the $^{111}In$-labelled paclitaxel as tumor imaging agent. In order to provide a taxol molecule with a functional group which is able to chelate In-111, taxol-DTPA conjugate and 2'-hemisuccinyltaxol were synthesized by esterification of taxol at C-2'on C-13 carbon with DTPA anhydride and succinic anhydride, respectively. Synthesis yield of the taxol derivatives was 34% for taxol-DTPA and 80% for 2'-hemisuccinyltaxol. Cytotoxicity of the taxol derivatives were measured by MTT method toward cell lines HT29, B16, P388, and CT26. The cytotoxic activities of the taxol derivatives were maintained, although less active than taxol. Radiolabelling of the taxol derivatives were proceeded directly with $^{111}InCl_3$ or indirectly with $^{111}In$-citrate(ligand-exchange method). The ligand-exchange method was not suitable because some precipitates appeared during the reaction. On the contrary, by direct radiolabelling method, we were able to obtain taxol-DTPA-$^{111}In$ in 100% radiochemical yield. However, 2'-hemisuccinyltaxol was not labelled by both methods. Yield and radiochemical purity of the radiolabelled com-pound were determined by HPLC, paper chromatography and instant thin layer chromatography. Taxol-DTPA-$^{111}In$ was characterized to be hydrophilic by lipophilicity test, and nearly non-adhesive to HT29, B16, P388, and CT26 by cell binding affinity test. Binding affinity of the taxol-DTPA-$^{111}In$ complex to serum proteins was also examined by protein precipitation with 30% trichloroacetic acid. The results showed that 30% of the taxol-DTPA-$^{111}In$ complex binds with serum proteins.

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Studies on the Cytotoxicity and Antimicrobial Effects of the Extract of Ajuga multiflora Bunge (조개나물 추출물의 세포독성과 항균효과)

  • Ryu, Myeng-Hwan;Aeam, Yong-Dea;Byun, Jong-Ho;Cho, Hoon;Yang, Eun-Yeong;Kang, Kil-Ung;Shin, Min-Kyo;Baek, Seung-Hwa
    • Korean Journal of Pharmacognosy
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    • v.31 no.1
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    • pp.72-76
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    • 2000
  • This study was carried out to evaluate cytotoxic effects of Ajuga multiflora Bunge extracts on murine leukemia tumor $(P388D_1)$ cell lines. Disruptions in cell organelles were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide (MTT) assay. The comparison of $IC_{50}$ values of Ajuga multiflora Bunge extracts in L1210 and $P388D_1$ cell lines showed that their susceptibility to these extracts decreased in the following order: Adriamycin>methanol extract>chloroform extract>ethyl acetate extract>hexane extract>water extract by the MTT assay. In order to develop an antimicrobial agent, dried Ajuga multiflora Bunge was extracted with several solvents, and then antimicrobial activity was investigated. The minimal inhibitory concentration (MIC) of the extract against microorganisms were also examined. Antimicrobial activities of amocla and ketoconazole as references were compared to those of extracts of $H_2O$, n-hexane, chloroform, ethyl acetate and methanol. The antimicrobial activity of all extracts from the sample had growth inhibition activity against gram-negative bacteria, gram-positive bacteria and fungi $(MIC>200\;{\mu}g/ml)$. These results suggest that the methanol soluble extract of Ajuga multiflora Bunge may be a valuable choice for the studies on the treatment of murine leukemia tumor cell lines.

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Screening for Antitumor Efficacy fro the wild plants in Korea(II) (韓國産 抗腫瘍性 資源의 Screening에 대하여(II))

  • Lee, Sang-Rae;Yoon, Eui-Soo;Shin, Soo-Cheol;Lee, Sang-Cheol
    • Korean Journal of Plant Resources
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    • v.6 no.1
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    • pp.25-32
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    • 1993
  • Many plants, which collected from Korea, were applied to antitumor and cytotoxic screeing tests against sarcom 180 a ascitec in mice, V-79 KB and P388 cultured cells. The results are summarixed as follows:1) The total packed cell volum method has been used for the antineoplastic screening for from natural higher plants in Korea. By this method, we have found out that the root, leaf and stem of Tripterygiu, regelii Spragne & Taketa having strong antineoplastic activity and also Rumex Japonicus Houtt. Eragrositis ferru-ginea Beauv. and Patrinia scabio-saefolia Fischer showed significant activity to anticancer tumor while cynanchum wilfordii Hemsley, and Rosa polyantha Sieb. et Zacc. showed slight activity to antitumor. 2) Among the 13 tested plants, the root and stem of Tripterygium regelii Spragne & Taketa and Amethystanthus excisus Nakai showed strong antitumor activity by the V79 cytotoxic cell screening test. 3) Twelve plants, which are glowing in mountainous area of Korea tested to anticancer activity. From the results, Eragrositis ferru-ginea Beauv., Angelica gigas Nakai, Geranium sibiricum L., Patrinia scabio-saefolia Fisher, Cynanchum wilfordii Hemsley, and Rubia akane Nakai have been proved to be anti-cancer plants by using P388 cell cultured method. 4) Tripterygiu, resgelii Spragne & Taketa, Eragrositis ferru-ginea Beauv., Patrinia scabio-saefoli Fisher, Cynanchum wilfordii Hemsley and Rasa polyantha Sieb. et Zacc., var. genuina Thunb. showed strong anti-tumor activity both total packed cell volume method and Cytotoxicity method.

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Effects of Aloe vera on the Cytotoxicity of Anticancer Drugs in Vitro (Aloe vera가 항암제의 세포독성에 미치는 영향)

  • 표명윤;윤지현
    • YAKHAK HOEJI
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    • v.43 no.1
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    • pp.104-110
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    • 1999
  • We investigated effects of methanol extract of Aloe vera on anticancer drugs(cisplatin, mitomycin C, 5-fluorouracil)-induced growth inhibition in p388, L1210, HCT-15, SK-HepG-1 as cancer cell lines and mouse splenocytes as a normal cell by MTT assay, respectively. We also examined the effects of aloe extract and mitomycin C on the mitogen(Con, A, LPS)-induced splenocyte proliferation. Aloe extract(0.25 mg/m , 1.25 mg/m , 2.5 mg/m , 5.0 mg/m ) showed dose-dependently selective cytotoxicity against the cancer cell lines. In contrast, Aloe extract increased the growth and proliferation of the normal mouse splenocytes. The combination of aloe extract with anticancer drugs showed an additive effect for the cytotoxicity against cancer cell lines. However, that combination reduced clealy the anticancer drugs-induced toxicity against the normal mouse splenocytes.

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Comparative Study on the Cytotoxic Activities of Red Ginseng of Korea and China (한국 및 중국 홍삼의 암세포 증식억제 효과 비교연구(III))

  • 황우익;손정원
    • Journal of Ginseng Research
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    • v.17 no.3
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    • pp.196-202
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    • 1993
  • A study was performed to compare the anticancer effects of Korean and Chinese red ginseng roots. The whole crude extracts or chloroform, methanol and acetone fractions of the crude extracts were added in the culture medium of three cancer cell lines, a mouse leukemia cell line ($P_{388}$), a human colon carcinoma cell line (HT-29) and a human rectal carcinoma cell line (HRT-18), to screen the growth inhibition effects. The results are summarized as follows : 1. Crude extracts of both Korean and Chinese red ginseng roots inhibited the proliferation of all the three cancer cell lines tested in a dose dependent manner. However, the growth inhibition effects of Korean red ginseng extracts were significantly greater than that of Chinese red ginseng. 2. An acetone fraction showed the greatest antiproliferative effects among the 11'hole crude extracts, chloroform, methanol and acetone fractions of the crude extracts. 3. These results suggest that the active antiproliferative components of the crude extracts are present mostly in the acetone fraction.

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Cytotoxic and Apoptotic Activites of Echinomycin Derivative (Echinomycin-7) on P388 Murine Leukemia Cells

  • Jeon, Hyang;Kim, Sung-Su;Kim, Yoon-Suk;Park, Yil-Sung;Kim, Yong-Hae;Choi, Sun-Ju;Kim, Soo-Kie;Kim, Tae-Ue
    • BMB Reports
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    • v.31 no.6
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    • pp.560-564
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    • 1998
  • Echinomycin-7 is an echinomycin derivative, Smethylated sulfonium perchlorate of echinomycin. We studied the in vitro cytotoxicity and in vivo antitumor activity of echinomycin-7 against P388 leukemia cells and compared the results with echinomycin. With respect to the cytotoxic effects, echinomycin-7 had cell line-dependent $IC_{50}$ values while echinomycin had similar values to several tumor cell lines. Also, in vivo antitumor activities were observed in tumor-bearing mice treated with both agents, which showed that echinomycin-7 had a broad therapeutic dose range. We also observed the apoptosis on leukemia cells treated with echinomycin-7 which exihibited the ladder pattern of DNA on electrophoresis. In addition to apoptosis, echinomycin-7 arrested $G_1/S$ phases of the cell cycle at the same time. We then examined the signaling pathway of echinomycin-7-induced apoptosis and showed that ERK of the MAP kinase family was activated and translocated into the nucleus by echinomycin-7 stimulation. This study suggests that echinomycin-7 acts as an antitumor agent through in vitro cytotoxicity and has in vivo antitumor activity against leukemia cells, and that the echinomycin-7- induced apoptosis might involve signal transduction via MAP kinases.

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Cytotoxicity and Structure Activity Relationship of Dammarane-Type Triterpenoids from the Bark of Aglaia elliptica against P-388 Murine Leukemia Cells

  • Hidayat, Ace Tatang;Farabi, Kindi;Harneti, Desi;Maharani, Rani;Darwati, Darwati;Nurlelasari, Nurlelasari;Mayanti, Tri;Setiawan, Arlette Suzy;Supratman, Unang;Shiono, Yoshihito
    • Natural Product Sciences
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    • v.23 no.4
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    • pp.291-298
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    • 2017
  • Six dammarane-type triterpenoids, dammar-24-en-$3{\beta}$-ol (1), $3{\beta}$-epicabraleahydroxy lactone (2), (E)-25-hydroperoxydammar-23-en-$3{\beta}$,20-diol (3), dammar-24-en-$3{\beta}$,20-diol (4), $3{\beta}$-acetyl-20S,24S-epoxy-25-hydroxydammarane (5), and $3{\beta}$-epiocotillol (6) were isolated from the methanolic extract of the bark of Aglaia elliptica. The chemical structure were identified on the basis of spectroscopic evidence and by comparison with those spectra previously reported. Compounds 1 - 6 were isolated first time from this plant. Compounds 1 - 6, along with a known synthetic analog, cabraleone (7) were evaluated their cytotoxic activity against P-388 murine leukimia cells in vitro. Among those compounds $3{\beta}$-acetyl-20S,24S-epoxy-25-hydroxydammarane (5) showed strongest cytotoxic activity with $IC_{50}$ value of $8.02{\pm}0.06{\mu}M$.