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Photomixotrophic Growth of Solanum tuberosum L. in vitro with Addition and Omission of Organic Materials at Thee Initial Sucrose Levels in the Medium (세 수준의 자당이 첨가된 배지에서 유기물의 첨가 유무에 따른 Solanum tuberosum L.의 기내 광혼합영양생장)

  • Jeong, Byoung-Ryong;Yang, Chan-Suk;Kim, Gyeong-Hee;Park, Young-Hoon;Kozai, Toyoki
    • Journal of Bio-Environment Control
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    • v.13 no.1
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    • pp.51-55
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    • 2004
  • The most commonly used inorganic nutrient compositions such as Murashige & Skoog medium have been optimized for heterotrophic growth. Therefore, they may not be optimal for photomixotrophic and photoautotrophic growth of plantlets. In photomixotrophic micropropagation, emdium sugar level is often lowered, while light and $CO_2$ levels in vessel are raised, and chlorophyllous explants are used to facilitate photosynthetic carbon acquisition. In a factorial experiment effect of addition (+) and omission(_) of organic materials (OM, 0.5 g ${\cdot}$ $m^{-3}$ each of thiamine, nicotinic acid and pyridoxine and 100 ${\cdot}$ $m^{-3}$ myo-inositiol) combined with three sucrose levels (0, 15, and 30 kg ${\cdot}$ $m^{-3}$) was tested on the growth of potato plantlets. Each of nodal cuttings with a leaf was cultured on 0.1${\times}$$10^{-4}m^{-3}$) MS agar ( 8 kg ${\cdot}$ $m^{-3}$) medium (pH 5.80 before autoclave) in glass test tubes (100 mm${\times}$25mm) capped with a sheet of transparent film with a 6 mm diameter gas permeable filter (5.1 air exchanges ${\cdot}$$h^{-1}$). Cultures were maintained in a room for 27 days at $23^{\circ}C$, 50% RH, 350-450${\mu}mol\;{\codt}\;mol^{-1}CO_2$, 16 h ${\cdot}$ $d^{-1}$ photoperiod at 13${\mu}mol\;{\codt}\;m\;{\codt}\;s^{-1}$ PPFD provided by white cool fluorescent lamps. Growth of potato plantlet in the +OM and -OM treatments were similar, while medium pH was 0.2 scale lower in the latter. Dry weight, % dry matter, and stem diameter enhanced, while shoot to root dry weight ratio, leaf area, chlorophyll concentration per gram dry weight, and medium pH decreased with increasing initial sucrose level. Interaction between OM and sucrose levels was observed in shoot length and medium pH. Results indicate that OM can be omitted from the medium without detrimental effect while addition of sucrose was beneficial for the photomixotrophic growth of potato plantlets under raised light and $CO_2$.

Production of Fungal Lipids - I. On Intracellular Fungal Lipids - (곰팡이 유지 생산에 관한 연구 - 제 1 보 : 곰팡이 세포내 유지에 대하여 -)

  • Yoo, Jin-Young;Shin, Dong-Hwa;Yim, Ho;Min, Byong-Yong;Suh, Kee-Bong
    • Korean Journal of Food Science and Technology
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    • v.12 no.2
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    • pp.97-102
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    • 1980
  • Cladosporium fulvum, Aspergillus ochraceus, Aspergillus terreus and N-1 (unidentified species) were cultured on the artificial media containing sucrose as a carbon source at 20, 25 and $30^{\circ}C$ for 10 to 12 days. The lipids in the felts were extracted with chloroform-methanol mixture and the class composition and fatty acids of the lipid were determined. The summarized results are as follows 1. The average felts produced by each species per 100 ml of media were $3.82{\pm}0.30g$ for Cl. fulvum, $2.62{\pm}0.23g$ for Asp. ochraceus, $4.24{\pm}0.25g$ for Asp. terreus and $4.62{\pm}0.10g$ for N-1. Their crude fat contents $27.5{\pm}1.61%,\;50.47{\pm}1.00%,\;46.6{\pm}1.59%$ and 33.78 % and the fat coefficient 6.92, 8.88, 13.01 and 10.28, respectively. 2. The lipids produced by these species were mainly composed of triglyceride and the next free fatty acid in Cl. fulvum and N-1 and phospholipid Asp. ochraceus and Asp. terreus. 3. The major fatty acids of the lipids were in order of oleic, palmitic, linoleic and stearic acids in Asp. ochraceus, Asp. terreus and Cl. fulvum and linoleic, palmitic, oleic and stearic acid in N-1. The total percentage contents of these major fatty acids were over 98 % the former and over 95 % the latter. 4. The constituent fatty acids of the lipid were changed depending on the incubation temperature but hardly found a certain tendency except linoleic acid which was higher at lower temperature. 5. The total percentages of unsaturated fatty acids in the lipids were $50{\sim}60%$ and comparatively higher at lower incubation temperature.

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The effect of neuropeptides on secretion of Interleukin-8(IL-8) (Interleukin-8 (IL-8) 분비에 미치는 neuropeptides의 영향에 관한 연구)

  • Kim, Kyung-Jun;Park, Sang-Hyuk;Choi, Kyoung-Kyu;Park, Sang-Jin
    • Restorative Dentistry and Endodontics
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    • v.31 no.3
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    • pp.153-160
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    • 2006
  • We investigated the secretion of Interleukin-8 (IL-8) from ginviva and periodontal ligament stimulated with Substance P (SP) and Calcitonin Gene-related Peptide (CGRP). Gingiva (GF), periodontal ligament (PDLF) and pu)p (PF) tissues were collected from extracted intact 3rd molars. Cultured cells were stimulated with different concentrations of SP for 4 hrs, and stimulated with SP, CGRP and Tumor Necrosis Factor-$\alpha$ (TNF-$\alpha$) for 8 hrs. Then RNase Protection Assay was carried out. ELISA was performed using supernatants of stimulated cells for quantitative analysis of IL-8. Results were assessed using student t-test with significance of P<0.05. According to this study, the results were as follows: 1. IL-8 mRNA was detected in all type of cells studied (PF, GF and PDLF) 2. IL-8 mRNA expression was not increased after stimulating 4 hrs with SP ($10^{-5}M$) and SP ($10^{-8}M$) compared with Mock stimulation in all type of cells studied. 3. IL-8 mRNA expression was not increased after stimulating 8 hrs with SP ($10^{-4}M$) and CGRP ($10^{-6}M$) compared with Mock stimulation in all type or cells studied. 4. TNF-$\alpha$ (2 ng/ml) increased the expression of IL-8 mRNA in all kind of cells studied. 5. The secretion of IL-8 from GF was increased 8 hrs after the stimulation with CGRP ($10^{-6}M$)(p<0.05). 6. The secretion of IL-8 from PDLF was. increased 8 hrs after the stimulation with SP ($10^{-4}M$)(p<0.05). Calcitonin Gene-related Peptide (CGRP) increased Interleukin-8 (IL-8) which plays an important role in chemotaxis of neutrophil in Calcitonin Gene-related Peptide (CGRP) gingival tissue , whereas Substance P increased the secretion of IL-8 from periodontal ligament.

The effect of tumor necrosis factor (TNF)-α to induce matrix metalloproteinase (MMPs) from the human dental pulp, gingival, and periodontal ligament cells (사람의 치수, 치은, 치주인대 세포에 tumor necrosis factor (TNF)-α로 자극 시 matrix metalloproteinase (MMPs)의 분비에 관한 연구)

  • Rhim, Eun-Mi;Park, Sang-Hyuk;Kim, Duck-Su;Kim, Sun-Young;Choi, Kyoung-Kyu;Choi, Gi-Woon
    • Restorative Dentistry and Endodontics
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    • v.36 no.1
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    • pp.26-36
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    • 2011
  • Objectives: In the present study, three kinds of tissues cells (pulp, gingiva, and periodontal ligament) were investigated if those cells express MMP and TIMP when they were stimulated with neuropeptides (substance P, CGRP) or proinflammatory cytokine, TNF-$\alpha$. Materials and Methods: The cells cultured from human dental pulp (PF), gingiva (GF) and periodontal ligament were (PDLF) stimulated with Mock, SP, TNF-$\alpha$, and CGRP for 24 hrs and 48 hrs. for an RNase protection assay and Enzyme Linked Immunosorbent Assay. Cells (PF, GF and PDLF) seeded in 100 mm culture dish were stimulated with SP ($10^{-5}$, $10^{-8}\;M$) or only with medium (Mock stimulation) for 4hrs and for 24 hrs for RNase Protection Assay, and they were stimulated with CGRP ($10^{-5}\;M$) and TNF-$\alpha$(2 ng/mL) for 24 hrs and with various concentraion of TNF-$\alpha$(2, 10, and 100 ng/mL) for Rnase Protection Assay with a human MMP-1 probe set including MMP 1, 2, 8, 7, 8, 9, 12, and TIMP 2, 3. In addition, cells (PF, GF and PDLF) were stimulated with Mock and various concentraion of TNF-$\alpha$(2, 10, and 100 ng/mL) for 24 hrs and with TNF-$\alpha$(10 ng/mL) for 48 hrs, and the supernatents from the cells were collected for Enzyme Linked Immunosorbent Assay (ELISA) for MMP-1 and MMP-13. Results: The expression of MMPs in PF, GF, PDLF after stimulation with SP and CGRP were not changed compared with Mock stimulation for 4 hrs and 24 hrs. The expression of MMP-1, -12, -13 24 hrs after stimulation with TNF-$\alpha$ were upregulated, however the expression of TIMP-3 in PF, GF, PDLF after stimulation with TNF-$\alpha$ were downregulated. TNF-$\alpha$(2 ng/mL, 10 ng/mL, 100 ng/mL) increased MMP-1 and MMP-12 expression in PF dose dependently for 24 hrs. Conclusions: TNF-$\alpha$ in the area of inflammation may play an important role in regulating the remodeling of dentin, cementum, and alveolar bone.

A Study on the Growth Environment and Tissue Culture of Gyrophora esculanta MIYOSHI in Korea (한국산(韓國産) 석이(石耳)의 생육환경(生育環境)과 조직배양(組織培養)에 관(關)한 연구(硏究))

  • Kim, Jai Saing
    • Journal of Korean Society of Forest Science
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    • v.78 no.4
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    • pp.333-344
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    • 1989
  • The objectives of this study were to investigate the growth of Gyrophora esculanta and to establish a method of tissue culture of the plant. The results obtained were as follows : 1. The Gyrophora esculanta was found growing mostly on the rock slopes of 722 m to 1915 min elevation on mountains in Korea. 2. Trees growing in the vicinity of the G. esculanta were mainly Quercus spp., Pinus thunbergii, Acer spp. and Lespedeza spp, Especially Quercus spp. was found growing in all of the study site. 3. The average Length of the rock slopes with G. esculanta growing on was 14 m and their aspects were mostly south. 4. The G. esculanta were found growing on rocks of Crystalline Schist, Quartz, Liparite, Granite, ete. Particularly they were mostly found on granites. The gradient of the rock slopes was in the range of 22-90 degrees. 5. The mean number of individuals of G. esculanta per one rock slope ranged from 14 at Mt. Bukhan to 70 at Mt. Jrri. Their mean diameter ranged from 1.8cm at Mt. Munsu to 4.6cm at Mt, Sokri. 6. The average percentage of G. esculanta with fruit body was 17.6%. The highest value was found at Mt. Cheonhwang (24.0%). 7. When the 100 segments of rhizoid of Gyrophora esculanta cultured in Detmer's medium supplemented with kinetine 5mg/l and 2, 4-D 3mg/l, n callus of microspore origins were induced from about 20% of the segments. As the induced n callus was transplanted on the six different types of rocks, it was observed that the juvenile G. esculanta grew best on granite and the development rate of G. esculanta on the granite was about 55%.

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A Study on the Cd Accumulation and Tolerance of Pisolithus Tinctorius in Vitro (배양기내(培養器內)에서 모래밭버섯균(菌)의 Cd 축적(蓄積)과 내성(耐性)에 관(關)한 연구(硏究))

  • Han, Sim Hee;Lee, Kyung Joon;Hyun, Jung Oh
    • Journal of Korean Society of Forest Science
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    • v.90 no.1
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    • pp.83-89
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    • 2001
  • This study was conducted to test the Cd accumulation and Cd-tolerance of Pisotithus tinctorius(Pt). Pt was isolated from Pinus thunbergii forest in Muan, Chonnam Province in 1997. Pt was cultured on MMN medium supplemented with $CdSO_4{\cdot}5H_2O$ at the final concentration of 0, 0.2, 0.5, 2, and $10{\mu}g/m{\ell}$ for 40 days. Growth rate and tolerance index of the fungus were measured every week, while Cd concentration, superoxide dismutase(SOD), and glutathione reductase(GR) of the fungus were analyzed at the end of the culturing, Pt showed growth reduction in vitro at $2{\mu}g/m{\ell}$ Cd in the medium and almost stopped growth at $10{\mu}g/m{\ell}$ Cd. Tolerance index of Pt decreased with increasing Cd concentration. Cd concentration of Pt was the highest at $10{\mu}g/m{\ell}$ Cd. Activities of SOD did not show significant difference between Cd concentrations, but GR of Pt increased at $0.5{\mu}g/m{\ell}$ Cd, and decreased at $2{\mu}g/m{\ell}$ Cd. Consequently Pt could be called Cd accumulator with a tolerance mechanism to Cd. Their tolerance to Cd were expressed through the higher production of antioxidants such as GR. Pt may be used for revegetation and decontamination of soil polluted by heavy metals.

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The Effect of EDTA and Fetal Cord Serum Supplementation to Ham's F-10 Culture Medium on Developmental Potential of Mouse Embryos In Vitro (Ham's F-10 배양액에 첨가된 태아제대혈청과 EDTA가 백서 수정란의 분할에 미치는 영향)

  • Kim, Byeong-Seog;Lee, Young-Gi;Park, Yoon-Kee;Lee, Tae-Hyung;Lee, Sung-Ho
    • Journal of Yeungnam Medical Science
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    • v.12 no.1
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    • pp.124-134
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    • 1995
  • It is the most important to select optimal culture conditions to promote safe embryo growth in the technique of human in vitro fertilization and embryo transfer. It has been shown that the addition of biologic fluids, such as blood serum, of various origins, improved fertilization and early cleavage rates in numerous species. The purpose of this study is to attempt to measure developmental potential of mouse eggs fertilized and cleaved in Ham's F10 culture medium containing a chelating agent, EDTA and fetal cord serum. In this study, we selected 40 female mice and 20 male mice, and investigated optimal serum concentrations for mouse embryo growth. Two cell stage mouse embryos were cultured in Ham's F-10 medium, Ham's F-10 medium with various concentrations of EDTA, or Ham's F-10 medium with EDTA and 10% human cord serum. Developmental ratios to morula in Ham's F-10 medium containing various concentrations of EDTA and/or 10% fetal cord serum were significantly higher than in unsupplemented Ham's F-10 medium (p<0.05). Developmental ratios to blastocyst in Ham's F-10 containing 10% fetal cord serum and $50{\mu}M$ or $100{\mu}M$ EDTA were significanltly higher than in unsupplemented Ham's F-10 medium (p<0.05). Developmental ratios to morula in Ham's F-10 containing 10% fetal cord serum and $100{\mu}M$ EDTA were significanltly higher than in Ham's F-10 with 10% fetal cord serum used commonly in many human IVF centers(p<0.05). Developmental ratio to blastocyst in Ham's F-10 containing 10% fetal cord serum and $100{\mu}M$ EDTA was significanlty higher than in Ham's F-10 with $200{\mu}M$ EDTA(P<0.05). In summary, embryo development to morula and blastocyst was significanlty higher in the presence of human cord serum or EDTA than in the unsupplemented medium. The most significanly development to morula and blastocyst was obtained at Ham's F-10 medium with $100{\mu}M$ concentration of EDTA and 10% fetal cord serum. These results suggest that Ham's F-10 medium containing 10% fetal cord serum and optimal concentrations of EDTA significantly promoted early cleavage of mouse zygotes, and these will be useful as basic data for the selection of culture medium in human in vitro fertilization.

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Cryopreservation of Umbilical Cord as a Source of Mesenchymal Stromal Cells and Growth Factors (간엽줄기세포와 성장인자의 공급원으로서 제대 조직의 동결 보관)

  • Lee, Hye Ryun;Roh, Eun Youn;Shin, Sue;Yoon, Jong Hyun;Kim, Byoung Jae;Jeon, Hye Won
    • The Korean Journal of Blood Transfusion
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    • v.23 no.2
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    • pp.115-126
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    • 2012
  • Background: Umbilical cord (UC) is a promising source of mesenchymal stromal cells (MSCs). We compared the characteristics of MSCs from cryopreserved UC with those from fresh tissues, and demonstrated the possibility of UC cryopreservation for acquisition of MSCs from cryopreserved UC. Methods: Each UC was sliced into two types ($1{\sim}2mm^3$ vs. 0.5 cm), and cryopreserved in liquid nitrogen using different media (autologous cord blood plasma, aCBP vs. RPMI 1640). A fresh aliquot of $1{\sim}2mm^3$-sized UC was used as control tissue. After one week, the cryopreserved tissues were thawed and cultured. For the 0.5 cm UC, a slicing step into $1{\sim}2mm^3$ was needed. Cell count, viability, proliferative activity, and surface antigens were determined from harvested MSCs. Several growth factors (EGF, IGF-1, PDGF, TGF-${\beta}$, bFGF, and VEGF), were measured from the culture supernatant. Results: Eleven UC were enrolled in the study. Efficiencies of obtaining MSCs were higher in cryopreserved UC using RPMI 1640, compared with use of aCBP; the same result was observed for 0.5 cm sized UC, compared with $1{\sim}2mm^3$ sized UC. No difference in proliferative activity was observed between MSCs from fresh and cryopreserved UC. The amount of growth factors in culture supernatant using RPMI 1640 was larger than that of fresh tissues. Conclusion: We obtained growth factors from the supernatant as well as MSCs from cryopreserved UC. As with a cord blood bank, in the future, cryopreservation of UC for acquisition of both MSCs and growth factors would be possible in a time of need.

Growth and Flowering of Campanula Species as Affected by Duration, Temperature, and Light Condition during Chilling Treatment (저온처리 기간, 온도 및 광 조건이 자생초롱꽃의 생육과 개화에 미치는 영향)

  • Lee, Young Mi;Park, Yoo Gyeong;Jeong, Byoung Ryong
    • FLOWER RESEARCH JOURNAL
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    • v.19 no.1
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    • pp.22-29
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    • 2011
  • The experiment investigated effect of duration, temperature, and light condition during chilling treatment on growth and flowering of four Campanula species in a factorial experiment. Two parent species, Campanula punctata Lam. var. rubriflora Mak. and C. Punctata Lam., and their two $F_1$ hybrids, C. punctata Lam. ${\times}$ C. punctata Lam. var. rubriflora Mak. ('Jiknyeo') and C. punctata Lam. var. rubriflora Mak. ${\times}$ C. punctata Lam. ('Gyeonu'), were used. Plants were cultured in vitro for five weeks at $25^{\circ}C$ under about $75{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ PPFD before being chilled at 4 or $25^{\circ}C$ for 3, 6, or 9 weeks under a darkened or lighted (about $10{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ PPFD) condition. After chilling treatment, plants were transplanted to 10 cm pots filled with a commercial growing medium and were transferred to environment-controlled growth chambers and subsequently to a greenhouse to observe their reproductive growth. Growth of all species and flowering of a $F_1$ hybrid 'Jiknyeo' were affected by duration, temperature, and light condition during chilling treatment. The greatest growth and survival percentage were observed in C. punctata Lam. var. rubriflora Mak. The survival percentage was greater when plants were chilled in a lighted than darkened condition, whereas it decreased when plants were chilled more than six weeks in vitro. Among the four species tested, flowering was observed only in a $F_1$ hybrid 'Jiknyeo' with 62.5% flowering plants when it was chilled at $25^{\circ}C$ for three weeks under a lighted condition. Percent flowering plant was affected by duration, temperature, and light condition during chilling treatment. Three-week chilling at $4^{\circ}C$ under a darkened condition significantly reduced days to flowering. These results suggest that the low temperature requirement for flowering is not qualitative but quantitative in Campanula species. Further experiment with more number of plants is necessary to ascertain this conclusion.

Induction of Apoptosis in HT-29 Human Colorectal Cancer by Aloin (인간 대장암 세포 HT-29에서 Aloin에 의한 Apoptosis 유도)

  • Yoo, Eun-Seon;Woo, Joong-Seok;Kim, Sung-Hyun;Lee, Jae-Han;Han, So-Hee;Jung, Soo-Hyun;Park, Young-Seok;Kim, Byeong-Soo;Kim, Sang-Ki;Park, Byung-Kwon;Jung, Ji-Youn
    • Journal of Food Hygiene and Safety
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    • v.34 no.5
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    • pp.495-501
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    • 2019
  • Aloin [1,8-Dihydroxy-10-(${\beta}$-D-glucopyranosyl)-3-(hydroxymethyl)-9(10H)-anthracenone], is a natural anthraquinone from aloe. It has been shown to have antioxidant and anticancer effects in various types of human cancer cells, but the anticancer effects of aloin in human colorectal cancer cells HT-29 have not been elucidated. In this study, possible mechanisms by which aloin exerts its apoptotic action in cultured human colorectal cancer HT-29 cells were investigated. The results of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay shows that treatment with aloin (0, 100, 200, 300 and $400{\mu}M$) reduced cell viability in a concentration-dependent manner in HT-29 and showed no effects on cell proliferation in A375SM and AGS cells. In addition, it was confirmed that apoptotic body was significantly increased as shown by 4',6-diamidino-2-phenylindole (DAPI) staining, and increased apoptosis rate by flow cytometry in HT-29 cells treated with aloin (0, 200 and $400{\mu}M$). We confirmed by western blotting that aloin activated Bax (pro-apoptotic), cleaved-poly (ADP-ribose) polymerase (PARP) and caspase-3, -8 and Bcl-2 (anti-apoptotic) were not changed compared with the control. Aloin induced up-regulation of phospho-p38 and down-regulation of phospho-extracellular signal-regulated kinase (ERK)1/2. Therefore, aloin suppressed the growth inhibitory effects by the induction of apoptosis in human colorectal cancer cells and has potential as a cancer preventive medicine.