• Parasites, Hosts and Diseases
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• 제29권2호
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• pp.121-128
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• 1991

Toxoplasma gondii를 HL-60 세포와 함께 in vitro 배양시 Toxopsasma 침투 정도가 각 세포에서 균일하지 않으므로 세포의 주기에 따른 일정 phase가 그 침투에 좋은 환경을 제공할 것이라는 추론으로 HL-60세포 주기를 동시화(synchronization)하여 각 stage에서 변화를 관찰하였다. 동시화는 과량의 thymidine이 DNA 합성을 억제함을 이용하여 2mM thymidine을 10시간 간격으로 각 24, 18시간 동안 처리하여 (double thymidine block method) S (synthetic) phase를 진행하는 세포를 얻었고 이후 30시 간 동안 13회의 간격을 두고 $5{\times}10^6/ml$의 Toxoplasma를 첨가하여 1시간 동안 배양하였다. 숙주세포의 동시화 정도는 (1) 3H-thymidine의 표지량 (2) mitotic index 측전 및 (3) 세포수의 증가를 통해 확인하였다. Toxoplasma의 침투 정도는 S phase중에서도 배지에서 thymidine을 제거한 후 3시간 경과시가 그 전후에 비해 6배 이상 높았으며 특히 이 시기는 DNA 합성이 최고가 되는 점과 일치하였다. 침투된 Toxoplasma 수의 변화 외에 세포의 모양도 상당한 변화가 있었고 이는 19시간 후 2번째 S phase에서도 약하나마 관찰되었다 실험 결과를 통해 특정 약 1시간 동안 일어나는 어떤 세포의 변화가 Toxoplasma 침투에 중요한 역할을 한다는 것을 알 수 있었다. 이는 원충 기생충의 숙주세포 흡착과 interiorization과정에 receptor가 관련되고 몇 receptor는 세포주기에 따라 발현이 조절되는 사실로부터 $G_1/S$ 경계부터 3시간째에 발현되면서 Togopzasma를 유인하는 receptor molecule의 존재 가능성을 시사하였다.

• 약학회지
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• 제40권2호
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• pp.218-224
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• 1996

This laboratory has recently reported the synthesis and in vitro antitumor activity of PT(II) complexes containing ethylenediamine and diphosphine. In view of the reports of others, cisplatin is toxic to the kidney since the kidney's vulnerability to PT(II) complexes may originate in its ability to accumulate and retain platinum to a greater degree than other organs. The in vitro cytotoxicity of these synthetic PT(II) complexes on the primary cultured proximal tubular cells of rabbit kidney and renal cortical cells of human kidney was investigated. Three endpoints for cytotoxicity tests were evaluated:3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), $^3H$-thymidine uptake and the glucose consumption tests. The rank order of sensitivity exhibited $^3H$-thymidine uptake>MTT>glucose consumption test. The agents with diphosphine leaving group were significantly less cytotoxic than cisplatin. Moreover, 1,2-bis(diphenylphosphino)ethane (DPPE) exhibited less cytotoxicity than 1.3-bis (diphenylphosphino)propane (DPPP) against on rabbit and human cultured kidney cells. Based on these results, the decreased nephrotoxicity of these new complexes over cisplatin appeared to be partially attributable to a leaving group of DPPP and DPPE. This novel class of platinum compound represents a valuable lead in the development of a "third-generation" agent.

• Bulletin of the Korean Chemical Society
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• 제25권7호
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• pp.1003-1008
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• 2004

A novel class of alkoxybenzylcyanoguanidine analogs as the inhibitors of restenosis was discovered, which showed the inhibitory effects on angiotensin II-induced cell proliferation, determined by $[^3H]$thymidine incorporation method. The compound, N'-(4-nitrophenyl)guanidine analog 19, showed 62% inhibition of $[^3H]$thymidine incorporation at 1 ${\mu}M$ concentration. In addition, the compound 19 inhibited intimal thickening dose-dependently after balloon injury, which suggests the therapeutic potential for restenosis.

• 약학회지
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• 제36권5호
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• pp.449-454
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• 1992

The effect of radioprotective ginseng protein fraction on DNA repair capacity was determined by measuring the amount of $^{3}H-thymidine$ incorporated into DNA in the process of repair synthesis for UV damaged DNA. CHO-Kl cells were prepared whose semiconservative replication was inhibited by trimethylpsoralen plus near-UV(PUVA) treatment. When the cells were exposed to UV light alone, the DNA repair capacity was increased at first and then decreased as UV dose increased. However, when the ginseng fraction was treated to the cells, the DNA repair capacity was kept increasing regardless of UV dose increment. When the concentration of protein contained in the added fraction was increased gradually, the repair capacity was also increased almost linearly showing dose-response relationship of the effect. These results suggest that the enhancement of DNA repair capacity of the cell can be one of the mechanisms of radioprotection by the ginseng fraction.

• 대한의생명과학회지
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• 제12권3호
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• pp.249-254
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• 2006

It has been reported that liver is a very important organ to xenotransplantation. Pig is known to be a most suitable species in transplantation of human organs. However, the physiological function of pig hepatocytes is not clear elucidated. Epidermal growth factor (EGF) is known to be a mitogen in various cell systems. Thus, we examined the effect of EGF on cell proliferation and its related signal cascades in primary cultured pig hepatocytes. EGF stimulates cell proliferation in a dose (>1ng/ml) dependent manner. EGF-induced increase of $[^3H]-thymidine$ incorporation was blocked by AG 1478 ($10^{-6}M$, an EGF receptor antagonist) genistein and herbymycin A (tyrosine kinase inhibitors, $10^{-6}M$), suggesting the role of activation and tyrosine phosphorylation of EGF receptor. In addition, EGF-induced increase of $[^3H]-thymidine$ incorporation was prevented by neomycin $(10^{-4}M)$, U73122 $(10^{-5}M)$ (phospholipase C [PLC] inhibitors), staurosporine ($(10^{-8}M)$, or bisindolylmaleimide I $(10^{-6}M)$ (protein kinase C [PKC] inhibitors), suggesting the role of PLC and PKC. Moreover, EGF-induced increase of $[^3H]-thymidine$ incorporation was blocked by PD 98059 (a p44/42 mitogen activated protein kinase [MAPK] inhibitor), SB 203580 (a p38 MAPK inhibitor), and SP 600125 (a JNK inhibitor). EGF increased the translocation of PKC from cytosol to membrane fraction and activated p42/44 MAPK, p38 MAPK and JNK. In conclusion, EGF stimulates cell proliferation via PKC and MAPK in cultured pig hepatocytes.

• 대한치과교정학회지
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• 제22권1호
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• pp.147-158
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• 1992

This study was aimed to compare the relative cytotoxicity of the five common orthodontic bonding materials (Concise. Mon-lok, Ortho-One, Super C, Transbond) using cell culture technique. DNA synthesis of the fibroblasts was assessed by $^3H$-thymidine uptake to evaluate the effect of the bonding materials on the growth of the cells. The human gingival fibroblasts were explanted from the buccal gingiva of 10 year-old girl and cultured in $\alpha$-MEM/10% FBS/1% antibiotics medium, $37^{\circ}C$, 5% $CO_2$ incuvator. The gingival fibroblasts were tested with the medium into which the bonding materials had been soaked for 1 week. Or the bonding materials were placed on the cells immediately or 2 weeks after polymerization. After 22 hours, $^3H$-thymidine was added into the microtest wells and after 24 hours, the uptake of $^3H$-thymidine was determined by liquid scintilation counter. The results of this study were as follows. 1. DNA synthesis was significantly decreased with Super C and Transbond than Ortho-One, when treated with medium into which the bonding materials had been soaked for 1 week. 2. DNA synthesis was significantly decreased with Concise, Super C and Transbond than control, when treated immediately after polymerization. 3. DNA syntehsis was significantly decreased with Concise, Super C and Transbond than Ortho-One, when immediately after polymerization. 4. There was no significant difference in DNA synthesis between the bonding materials, when treated 2 weeks after polymerization.

• 동의생리병리학회지
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• 제19권3호
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• pp.623-627
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• 2005

Abnormal thyroid cell proliferation has a very important role in hypothyroidism. Thyroid stimulating hormone (TSH) stimulates proliferation and maintains differentiated function in thyroid follicular cells. A functioning rat thyroid cell line (FRTL) was used to study the effect of Gamgung-tang (GGT, Glycyrrhizae Radix, black beans, Angelicae Radix and Cnidii Rhizoma) on proliferation and cAMP accumulation of thyrocytes. Proliferation of cell was assessed by DNA synthesis and incorporation of $[^3H]thymidine$ into DNA. The concentration of cAMP was measured simultaneously with growth assessment. Extract of GGT ($0.15{\sim}0.9\;mg/ml$ increased DNA synthesis in a dose-dependent manner. GGT at 0.6 (p<0.05) and 0.9 mg/ml (p<0.01) significantly increased $[^3H]thymidine$ incorporation. A comparable effect was observed with TSH. GGT also enhanced cAMP accumulation. These results indicate that GGT increases the proliferation of thyrocytes and may be considered a promising agent for the treatment of autoimmune thyroid disease.

• Applied Microscopy
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• 제39권3호
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• pp.205-218
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• 2009

이 실험은 Ehrlich 종양세포를 이식한 후 BCG를 투여하였을 때, 위점막 상피세포의 형태학적 변화와 DNA합성능의 변화를 연구하고자 시행하였다. 실험동물로는 체중 25 g 내외의 성숙한 생쥐(ICR계통)를 정상대조군, 종양세포이식대조군(종양대조군), 종양세포이식후 BCG투여군(BCG투여군)으로 구분하였다. 종양대조군과 BCG투여군 동물들은 샅부위 피하에 각각 $1{\times}10^7$의 Ehrlich 종양세포를 이식한 후, 다음날부터 BCG ($0.6{\times}10^8{\sim}6.4{\times}10^8$ CFU, 27 mg/vial, Connaught Lab., Canada)를 하루건너 한 번씩 피부밑조직에 주사하였으며, 종양대조군은 종양세포이식 후에 BCG 대신 0.2mL의 생리식염수를 피부밑조직에 주사하였다. 자기방사법적 관찰을 위해서는 BCG를 마지막으로 주사한 다음날 $^3H$-thymidine (methyl-$^3H$-thymidine: specific activity 25 Ci/mmol, Amersham Lab., England) 0.7${\mu}Ci/gm$를 꼬리정맥에 주사하고, 70분 후 도살하여 위조직을 떼어내어 10% formalin에 고정하였다. 자기방사표본관찰은 위점막 조직이 세로로 잘 절단된 부위를 택하여 점막근육판을 따라 점막길이 3.5mm의 위점막 조직에 분포하는 $^3H$-thymidine 표지세포의 수를 계수하였으며, 일반조직 관찰을 위해서는 hematoxylin-eosin (H-E)염색을 시행하였다. 전자현미경 관찰을 위해서는 떼어낸 위조직을 2.5% glutaraldehyde-1.5% paraformaldehyde 혼합액에 고정한 후, 1% osmium tetroxide 액에 다시 고정하였으며, 고정이 끝난 조직은 탈수과정을 거쳐 araldite 혼합액에 포매하였다. 광학현미경적 관찰에서 종양대조군과 BCG투여군은 위점막 조직에서 형태적으로 큰 변화를 볼 수 없었다. 전자현미경적 관찰에서 BCG투여군의 점액상피세포는 전체적인 모습이 정상대조군과 종양대조군의 소견과 유사하였으나 정상대조군과 종양대조군에 비해 수초구조와 뭇소포체(multivesicular body) 및 전자밀도가 높은 큰 미토콘드리아속과립이 자주 관찰되었다. 자기방사법적 관찰에서 정상대조군, 종양대조군, BCG투여군은 점막길이 3.5 mm 당 출현하는 표지세포수가 각각 380.2 (${\pm}31.35$), 426.1 (${\pm}28.43$) 및 301.8 (${\pm}34.63$)개이었으며, BCG투여군은 정상대조군과 종양대조군에 비하여 표지된 은입자의 수가 매우 적어서 표지과립이 겨우 구별될 정도의 세포가 많이 관찰되었다. 이상의 결과를 종합해보면 Ehrlich 종양세포를 이식한 동물에 BCG를 반복 투여하면 위점막 상피세포의 DNA합성을 효과적으로 억제하면서도 형태적인 변화가 매우 경미하였으며, 이러한 결과는 BCG가 항암치료 시 보조제로 사용할 수 있는 좋은 약제라고 생각된다.

• 대한핵의학회지
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• 제19권1호
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• pp.83-93
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• 1985

The ability of $^{67}Ga$, administered carrier free as the citrate complex, to localize in human and animal tumors to an extent sufficient to permit visualization of the lesion by scanning is well established. However, neither the mechanism of $^{67}Ga$ uptake by tumors or inflammatory cells nor its relationship to cell type or to the biochemical status of the cell is yet understood. Author investigated the uptake of $^{67}Ga-citrate$ using subcellular tissue fractionation of rat livers treated with $CCl_4$ associated with the $^3H-thymidine$ incorporation rate to detect subcellular localization of $^{67}Ga$ and it's relationship in DNA synthesis. Large amounts of $^{67}Ga$ associated with the soluble portion of tissue homogenate rather than with isolated cell organelles and not related nuclei residue in the regenerating period after hepatocellular injury caused by $CCl_4$. The elevated uptake of $^{67}Ga$ in the livers of $CCl_4$ treated rats was also inhibited when protein synthesis was stopped by cyclohexamide. Thus protein and the soluble portion of issue homogenates seems to play an important role in the elevated uptake of $^{67}Ga$ in liver injury induced by $CCl_4$ treated rats.

• Archives of Pharmacal Research
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• 제20권3호
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• pp.259-263
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• 1997

In the preparation of acyclic thymidine nucleoside analogues, $K_2CO_3$(or NaH) treated thymine in DMSO was alkylated with .omega.-chloroalkyl nitrite (Cl-(CH_2)n-CN; n=1, 2, 3, 4) to provide an isomeric mixture of 1-(${\omega}$-cyanoalkyl)thymine (2a-d) and 1,3-bis(${\omega}$-cyanoalkyl)thymine in approximately 5:1 ratios. Reduction of the cyano function 2a-d with $NaBH_{4}/CoCl_{2}$ center dot$ 6H_{2}O$gave the corresponding 1-(${\omega}$aminoalkyl)thymine (3a-d). The newly formed primary amino function in 3a-d was directly reacted with 2-chloroethylisocyanate to afford the 1-[.omega.($N^{I}$-2-chloroethylureido) alkyl]thymine (4a-d) in good yields. Nitrosation of 1-[5-($ N^{I}-2$-chloroethylureido)pentyl] thymine (4d) with glacial acetic acid and dry $NaNO_{2}$-powder in anhydrous $CH_{2}Cl_{2}$gave two types of regioisomeric nitrosoureas, 1-[5-($N^{I}$--chloroethyl-$N^{I}$--nitrosoureido)pentylithymine (5d) and 1-[5-($N^{I}-2$-chloroethyl-N-nitrosoureido)pentyllthymine in approximately 5 :1 ratios. The in vitro cytotoxicity of the synthesized compounds (2a-d, 3a-d, 4a-d and 5a-d) against three cell lines (K-562, P-388 and FM-3A) are measured as $IC^{50}$ values. Compounds 3b and 4c showed moderate activities against all three cell lines, and all other compounds were found to be not active.