• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• 제27권4호
• /
• pp.314-320
• /
• 2001

Matrix metalloproteinases have long been viewed as ideal candidates for proteinases that enables tumor cells to permeated basement membrane defenses and invade surrounding tissue. There is growing evidence that the MMPs have an expanded role, as they are important for the creation and maintenance of a microenvironment that facilitates growth and angiogenesis of tumors at primary and metastatic sites. MT-MMPs are not secreted but instead remaining attached to cell surfaces. Although not all of the MT-MMPs are fully characterized, MT-MMPs have important role in localizing and activating secreted MMPs. The MMP genes are transcriptionally responsive to a wide variety of oncogene, growth factors, cytokine, and hormones. Currently, a number of MMP inhibitors are being developed and some have reached clinical trials as anti-metastatic or anti-cancer therapies. MT1-MMP is involved in the activation of proMMP-2. MT1-MMP is significant not only as a tumor marker but as a new target for chemotherapy against cancer. The purpose of this study was to evaluate the effects of protein kinase C inhibitor(genistein) on the proliferation of HT1080 and expression of MT1-MMP mRNA. Human fibrosarcoma cell line HT1080 was cultured and divided 2 groups. The experimental group was treated with $100{\mu}M$ genistein and incubated 12h, 24h for $[3^H]-thymidine$ uptake assay and northern hybridization individually. And the control group was treated with same amount of PBS for the above procedures. $[3^H]-thymidine$ incorporation was measured with ${\beta}$ ray detector. And RT-PCR and northern blotting for MT1-MMP mRNA was performed. The results were as follows 1. $[3^H]-thymidine$ uptake was reduced in experimental group with statistical significance. 2. MT1-MMP mRNA expression was significantly reduced in experimental group. These results showed that protein kinase C inhibitor (genistein) inhibited proliferation of HT1080 and almost completely blocked transcription of MT1-MMP mRNA. So, it is possible to use the protein kinase inhibitor (genistein) as anti-metastatic and anti-proliferative agent.

• 대한치과교정학회지
• /
• 제27권6호
• /
• pp.929-941
• /
• 1997

가장 바람직한 교정력은 환자에게 불편감을 주지 않고 치조골 상실과 치근흡수와 같은 조직의 손상없이 가장 빨리 치아를 이동시키는 힘이다. 최적의 교정력을 얻기 위하여 그 동안 많은 방법들이 시도되어 왔으며 최근에는 자석의 사용이 고려되고 있다. 본 연구는 Sm-Co 자석의 정자기장이 효소와 세포 활성에 미치는 영향을 알아보기 위하여 시행되었다. 적혈구 침강속도가 측정되었으며, 철이온과 관련된 효소 (Catalase, NO synthase)와 철이온과 무관한 효소 (Lactic dehydrogenase)의 활성과 세포내 합성은 Spectrophotometer를 이용하여 측정되었으며, 조골세포 $MC_{3}T_3-E_1$의 성장과 증식은 Crystal violet 염색법과 ${^3}H$-thymidine incorporation에 의한 DNA합성능을 측정하였다. 실험군의 적혈구는 표면자기장이 1,400 G (gauss)인 자석에, 효소와 조골세포는 7,000 G의 정자기장에 노출시키고, 정자기장에 노출시키지 않은 경우와 비교하여 다음과 같은 결과를 얻었다. 1. 적혈구 침강속도는 정자기장의 영향을 받지 않았다. 2. Catalase와 Lactic dehydrogenase의 활성은 정자기장의 영향을 받지 않았다. 3. NO synthase와 Lactic dehydrogenase의 세포내 합성은 정자기장의 영향을 받지 않았다. 4. 세포배양된 조골세포 $MC_{3}T_3-E_1$의 성장과 증식은 정자기장의 영향을 받지 않았다. 이상의 결과로 보아 정자기장은 효소와 세포 활성에 대한 영향이 없는 것으로 사료되었다.

• 대한치과교정학회지
• /
• 제21권1호
• /
• pp.97-111
• /
• 1991

It is the aim of this study to investigate the effects of sodium fluoride and sodium orthovanadate upon the proliferation and activity of the osteoblast (MC3T3-E1 cells). MC3T3-E1 cells were cultured in $\alpha-MEM$ containing $10\%$ FBS and various concentration of sodium fluoride and sodium orthovanadate was appended to serum free media. DNA synthesis was examined through the $[^3H]$ thymidine incorporation into DNA. Collagen synthesis was examined through the $[^3H]$ proline incorporation into collagenase digestible protein and noncollagen protein. The following results were drawn; 1. Sodium fluoride stimulated the DNA synthesis of osteoblast significantly in dose-dependent manner within the concentration from $2{\mu}M$ to $10{\mu}M$ (P < 0.005). 2. Sodium orthovanadate stimulated the DNA synthesis of osteoblast significantly in dose-dependent manner within the concentration from $2{\mu}M\;to\;8{\mu}M$, however showed diminution at $10{\mu}M$ (P < 0.001). 3. Sodium fluoride and sodium orthovanadate stimulated the percent collagen synthesis of osteoblast significantly in dose-dependent manner within the concentration from $5{\mu}M$ to $10{\mu}M$ (P < 0.001). 4. Sodium fluoride and sodium orthovanadate stimulated the noncollagen synthesis of osteoblast significantly in dose-dependent manner within the concentration from $5{\mu}M\;to\;10{\mu}M$ (P < 0.001). In conclusion, sodium fluoride and sodium orthovanadate stimulate the proliferation and activity of osteoblast by stimulation of DNA synthesis and collagen and noncollagen synthesis in osteoblast.

• 생약학회지
• /
• 제22권2호
• /
• pp.95-100
• /
• 1991

Salviae miltiorrhizae(SM) Radix extract increased $[^3H]-thymidine$ incorporation into rat hepatocytes at the concentration ranging from $5{\times}10^{-5}\;to\;5{\times}10^{-1}mg/ml$. It decreased the activities of s-GOT and s-GPT in cirrhotic rats induced by $CCl_4$, TAA and D-GalN, respectively and reduced the sleeping time induced by hexobarbital in $CCl_4$,TAA and D-GalN intoxicated mice, respectively. SM extract shortened the half-life of sulfobromophthalein in $CCl_4$ intoxicated rats.

• 대한미생물학회지
• /
• 제16권1호
• /
• pp.65-70
• /
• 1981

A microculture technique for the mixed lymphocyte culture test(MLC) in the sheep is described. The optimal incubation periods for the MLC are varied from 4 to 6 days depending on the sources of lymphocytes. It is found that a 2.5:1 stimulator-responder cell ratio and the responder cell concentration of $1{\times}10^5$ cells/well produce the highest($^3H$)-thymidine incorporation. Moreover cryopreservation of bovine lymphocytes results in satisfactory and reproducible MLC reaction. It is also evident that the MLC reactions of the sheep are under the control of histocompatibility matching between the stimulator cells and the responder cells. Significance of the technique as a too! for study on cell mediated immunity in sheep system, either normal or squamous cell carcinoma-bearing, are discussed.

• Journal of Plant Biology
• /
• 제36권1호
• /
• pp.19-27
• /
• 1993

Effects of polyamines on DNA synthesis were studied in synchronized culture of Nicotiana tabacum L. When DFMO and DFMA, inhibitors of ornithine decarboxylase and arginine decarboxylase, respectively were initially applied to the cells, the polyamine contents were rapidly dropped and [methyl-3H] thymidine incorporation into DNA was markedly reduced during the early stage of culture period. Inhibition of DNA synthesis, however, was partially reversed when these inhibitors were applied simultaneously with putrescine. In addition, exogenous administration of putrescine also increased the DNA synthesis during the all over the culture period. In vitro activity of DNA polymerase from Nicotiana tabacum L. was promoted by increasing concentrations of polyamines in the reaction mixture. Maximal activity was shown at 5 mM putrscine, 0.5 mM spermidine and spermine, respectively. Lack of Mg2+ ion in the reaction buffer resulted in an inhibition of the enzyme activity by about 30%. The inhibition could not be completely reversed by application of polyamines at optimal concentrations. These results suggest that polyamines promote the DNA synthesis in vivo and in vitro by stabilizing the DNA-helix upon binding to negatively charged groups on DNA or increasing the activity of DNA polymerase in Nicotiana tabacum L.

• 생약학회지
• /
• 제13권1호
• /
• pp.14-19
• /
• 1982

Potential toxicity of 15 medicinal plants used for herbal drugs, which were also described as being tonic for hematopoietic system or being toxic for the system in a oriental book 'Dong Ee Bo Gam', were evaluated in mice. Six plants among 15 plants tested appeared to exhibit acute toxicity along with bone marrow depression or with abnormally enhancing the $^3H-thymidine$ incorporation into DNA biosynthesis in bone marrow cells. Six plants were Paeonia albiflora, Pharbitis nil, Cemphalia lapidescens, Scutellaria baicalensis, Akebia quinata and Glycyrriza uralensis.

• 대한약학회:학술대회논문집
• /
• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
• /
• pp.108.2-108.2
• /
• 2003

Kaempferol, a flavonol compound, has been reported as the anti-oxidant and anti-angiogenic agent and it has been found to inhibit cell growth in vitro. Abnormal proliferation of vascular smooth muscle cells (VSMCs) plays an important role in development of atherosclerosis. In this study, we examined the anti-proliferative effect and its mechanism on rat aortic VSMCs treated by kaempferol. kaempferol significantly inhibited the platelet-derived growth factor (PDGF)-BB-induced proliferation of rat aortic VSMCs in concentration-dependent manner by cell count and ［$^3$H］-thymidine incorporation assay. (omitted)

• Clinical and Experimental Reproductive Medicine
• /
• 제33권2호
• /
• pp.115-123
• /
• 2006

목 적: 본 연구를 통해 GnRH 의한 세포 분열의 억제는 integrin, FAK 빛 ERK를 통한 세포 내 신호전달 기전을 통하여 일어남을 규명하고자 하였다. 연구방법: 연구에 사용된 인간자궁내막암 세포주는 DMEM/F12 (10% FBS)의 조건에서 배양 하였다. GnRH-I과 -II는 실험 목적에 따라 100 nM 농도로 0, 5, 10, 15, 20, 30분간 또는 10 nM or 100 nM의 농도로 20분간 처리 하였다. 세포의 분열 정도는 [$^3H$] thymidine incorporation assay를 이용하여 정량적으로 측정 하였으며, Immunoblotting 방법을 이용하여 단백질의 발현을 확인 하였다. 결 과: GnRH-I과 -II 모두 HEC1A 세포의 세보분열을 억제하였으며 integrin ${\beta}3$의 발현을 증가 시켰다. GnRH-I과 -II를 처리 후 FAK 및 ERK의 안산화가 증가됨을 관찰할 수 있었다. 결 론: GnRH에 의한 세포분열의 억제는 integrin의 발현과 FAK 및 ERK의 인산화 과정을 통하여 일어남을 알 수 있었다.

• Asian Pacific Journal of Cancer Prevention
• /
• 제13권11호
• /
• pp.5653-5657
• /
• 2012

Objectives: To investigate the effect of glycopeptide-preferring polypeptide GalNAc transferase 1 (ppGalNAc T1 ) targeted RNA interference (RNAi) on the growth and migration of human bladder carcinoma EJ cells in vitro and in vivo. Methods: DNA microarray assays were performed to determine ppGalNAc Ts(ppGalNAc T1-9) expression in human bladder cancer and normal bladder tissues. We transfected the EJ bladder cancer cell line with well-designed ppGalNAc T1 siRNA. Boyden chamber and Wound healing assays were used to investigate changes of shppGalNAc T1-EJ cell migration. Proliferation of shppGalNAc T1-EJ cells in vitro was assessed using [3H]-thymidine incorporation assay and soft agar colony formation assays. Subcutaneous bladder tumors in BALB/c nude mice were induced by inoculation of shppGalNAc T1-EJ cells and after inoculation diameters of tumors were measured every 5 days to determine gross tumor volumes. Results: ppGalNAc T1 mRNA in bladder cancer tissues was 11.2-fold higher than in normal bladder tissues. When ppGalNAc T1 expression in EJ cells was knocked down through transfection by pSUPER-shppGalNAc T1 vector, markedly reduced incorporation of [3H]-thymidine into DNA of EJ cells was observed at all time points compared with the empty vector transfected control cells. However, ppGalNAc T1 knockdown did not significantly inhibited cell migration (only 12.3%). Silenced ppGalNAc T1 expression significantly inhibited subcutaneous tumor growth compared with the control groups injected with empty vector transfected control cells. At the end of observation course (40 days), the inhibitory rate of cancerous growth for ppGalNAc T1 knockdown was 52.5%. Conclusion: ppGalNAc T1 might be a potential novel marker for human bladder cancer. Although ppGalNAc T1 knockdown caused no remarkable change in cell migration, silenced expression significantly inhibited proliferation and tumor growth of the bladder cancer EJ cell line.