• Title/Summary/Keyword: colony formation

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Knockdown of a Proliferation-inducing Ligand (PRIL) Suppresses the Proliferation of Gastric Cancer Cells

  • Cui, Jiu-Wei;Li, Yan;Wang, Chang;Yao, Cheng;Li, Wei
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.2
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    • pp.633-636
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    • 2012
  • Purpose: PRIL (proliferation-inducing ligand) is a newly identified member of the tumor necrosis factor (TNF) family and modulates death ligand-induced apoptosis. Here, we investigated the effect of PRIL on cellular characteristics relating to tumor progression in human gastric cancer. Method: Recombinant lentivirus containing PRIL siRNA was constructed and then infected MGC803 and SGC7901 gastric cancer cells. MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] colony formation and cell cycle analysis were used to study the effect of PRIL knockdown on gastric cancer cell proliferation. Results: PRIL expression in lentivirus infected cells was significantly reduced as evidenced by quantitative real-time PCR. Cell viability and colony formation of MGC803 and SGC7901 cells were significantly hampered in PRIL knock-down cells. Moreover, the cell cycle was arrested at G2/M phase, elucidating the mechanism underlying the inhibitory effect of siRNA on cell proliferation. Conclusions: Our study indicated that PRIL functions in promoting cell growth, and lentivirus-mediated PRIL gene knockdown might be a promising strategy in the treatment of gastric cancer.

miR-485 Acts as a Tumor Suppressor by Inhibiting Cell Growth and Migration in Breast Carcinoma T47D Cells

  • Anaya-Ruiz, Maricruz;Bandala, Cindy;Perez-Santos, Jose Luis Martin
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.6
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    • pp.3757-3760
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    • 2013
  • MicroRNAs (miRNAs) are small, non-coding RNAs (18-25 nucleotides) that post-transcriptionally modulate gene expression by negatively regulating the stability or translational efficiency of their target mRNAs. In this context, the present study aimed to evaluate the in vitro effects of miR-485 mimics in breast carcinoma T47D cells. Forty-eight hours after T47D cells were transfected with miR-485 mimics, an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was utilized to determine the effects on cell viability. Colony formation and cell migration assays were adopted to determine whether miR-485 affects the proliferation rates and cell migration of breast carcinoma T47D cells. Our results showed that ectopic expression of miR-485 resulted in a significant decrease in cell growth, cell colony formation, and cell migration. These findings suggest that miR-485 might play an important role in breast cancer by suppressing cell proliferation and migration.

Effects of Penicillin G on Morphology and Certain Physiological Parameters of Lactobacillus acidophilus ATCC 4356

  • Khaleghi, M.;Kermanshahi, R. Kasra;Zarkesh-Esfahani, S.H.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.8
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    • pp.822-829
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    • 2011
  • Evidence shows that probiotic bacteria can undergo substantial structural and morphological changes in response to environmental stresses, including antibiotics. Therefore, this study investigated the effects of penicillin G (0.015, 0.03, and 0.06 mg/l) on the morphology and adhesion of Lactobacillus acidophilus ATCC 4356, including the colony morphotype, biofilm production, hydrophobicity, $H_2_O2$ formation, S-layer structure, and slpA gene expression. Whereas only smooth colonies grew in the presence of penicillin, rough and smooth colony types were observed in the control group. L. acidophilus ATCC 4356 was found to be hydrophobic under normal conditions, yet its hydrophobicity decreased in the presence of the antibiotic. No biofilm was produced by the bacterium, despite testing a variety of different culture conditions; however, treatment with penicillin G (0.015-0.06 mg/l) significantly decreased its production of $H_2_O_2$ formation and altered the S-layer protein structure and slpA gene expression. The S-protein expression decreased with 0.015 mg/l penicillin G, yet increased with 0.03 and 0.06 mg/l penicillin G. In addition, the slpA gene expression decreased in the presence of 0.015 mg/l of the antibiotic. In conclusion, penicillin G was able to alter the S-layer protein production, slpA gene expression, and certain physicochemical properties of Lactobacillus acidophilus ATCC 4356.

miR-9 Modulates Osteosarcoma Cell Growth by Targeting the GCIP Tumor Suppressor

  • Zhu, Shao-Wen;Li, Jian-Peng;Ma, Xin-Long;Ma, Jian-Xiong;Yang, Yang;Chen, Yang;Liu, Wei
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.11
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    • pp.4509-4513
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    • 2015
  • Osteosarcoma is the most common primary bone tumor in humans, especially in childhood. However, the genetic etiology for its pathogenesis remains elusive. It is known that microRNAs (miRNAs) are involved in the development of tumor progression. Here we show that microRNA-9 (miR-9) is a potential oncogene upregulated in osteosarcoma cells. Knockdown of miR-9 in osteosarcoma resulted in suppressed colony formation and cell proliferation. Further study identified GCIP, a Grap2 and cyclin D interacting protein, as a direct target of miR-9. In addition, GCIP overexpression activated retinoblastoma 1 (Rb) and suppressed E2F transcriptional target expression in osteosarcoma cells. Moreover, GCIP depletion reversed miR-9 knockdown induced colony formation and cell proliferation suppression. In sum, these results highlight the importance of miR-9 as an oncogene in regulating the proliferation of osteosarcoma by directly targeting GCIP and may provide new insights into the pathogenesis of osteosarcoma.

Effects of the Developmental Stage of Extract Donor Embryos on the Culture of Marine Medaka Oryzias dancena Embryonic Stem Cell-like Cells (배아추출물 공여 배아의 발생단계가 바다송사리(Oryzias dancena) 배아 줄기세포 유사세포의 배양에 미치는 영향)

  • Ryu, Jun Hyung;Gong, Seung Pyo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.2
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    • pp.160-168
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    • 2017
  • Optimizing the conditions for stem cell culture is an essential prerequisite for the efficient utilization of stem cells. In the culture of fish embryonic stem cells (ESCs) or ESC-like cells, embryo extracts are important for stable growth, but there is no rule for determining the developmental stage of the embryos used to obtain extracts. Therefore, this study investigated the effects of the developmental stage of extract donor embryos on the culture of Oryzias dancena ESC-like cells. O. dancena ESC-like cells were cultured in different media containing each of four types of embryo extract depending on the developmental stage of the extract donor embryos. Growth, morphology, colony-forming ability, alkaline phosphatase (AP) activity, and embryoid body (EB) formation of the cells were investigated. While the developmental stage of the extract donor embryos did not influence the growth, morphology, AP activity, or EB formation of ESC-like cells, colony-forming ability was affected and the pattern of the effects differed completely between the two ESC-like cells investigated. These results suggest that the developmental stage of extract donor embryos should be selected carefully for the culture of ESC-like cells, according to the research purpose and type of cell line.

The Synergistic Anticancer Effect of Artesunate Combined with Allicin in Osteosarcoma Cell Line in Vitro and in Vivo

  • Jiang, Wei;Huang, Yong;Wang, Jing-Peng;Yu, Xiao-Yun;Zhang, Lin-Yi
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.8
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    • pp.4615-4619
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    • 2013
  • Background: Artesunate, extracted from Artemisia annua, has been proven to have anti-cancer potential. Allicin, diallyl thiosulfinate, the main biologically active compound derived from garlic, is also of interest in cancer treatment research. This object of this report was to document synergistic effects of artesunate combined with allicin on osteosarcoma cell lines in vitro and in vivo. Methods: After treatment with artesunate and allicin at various concentrations, the viability of osteosarcoma cells was analyzed by MTT method, with assessment of invasion and motility, colony formation and apoptosis. Western Blotting was performed to determine the expression of caspase-3/9, and activity was also detected after drug treatment. Moreover, in a nude mouse model established with orthotopic xenograft tumors, tumor weight and volume were monitored after drug administration via the intraperitoneal (i.p.) route. Results: The viability of osteosarcoma cells in the combination group was significantly decreased in a concentration and time dependent manner; moreover, invasion, motility and colony formation ability were significantly suppressed and the apoptotic rate was significantly increased through caspase-3/9 expression and activity enhancement in the combination group. Furthermore, suppression of tumor growth was evident in vivo. Conclusion: Our results indicated that artesunate and allicin in combination exert synergistic effects on osteosarcoma cell proliferation and apoptosis.

Efficient Production of Parthenogenetic Murine Embryonic Stem Cells by the Treatment of Pluripotin (SC-1) (Pluripotin(SC-1) 처리를 통한 단위발생 마우스 배아줄기세포 생산 효율 향상)

  • Kang, Hoin;Roh, Sangho
    • Journal of Embryo Transfer
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    • v.27 no.3
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    • pp.171-174
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    • 2012
  • Various small molecules can be used to control major signaling pathways to enhance stemness and inhibit differentiation in murine embryonic stem cell (mESC) culture. Small molecules inhibiting the fibroblast growth factor (FGF)/ERK pathway can preserve pluripotent cells from stimulation of differentiation. In this study, we aimed to evaluate the effect of pluripotin (SC-1), an inhibitor of the FGF/ERK pathway, on the colony formation of outgrowing presumptive mESCs. After plating the zona pellucida-free blastocyst on the feeder layer, attached cell clumps was cultured with SC-1 until the endpoint of the experiment at passage 10. In this experiment, when the number of colonies was counted at passage 3, SC-1-treated group showed 3.4 fold more mESC colonies when compared with control group. However, after passage 4, there was no stimulating effect of SC-1 on the colony formation. In conclusion, SC-1 treatment can be used to promote mESC generation by increasing the number of early mESC colonies.

Immunostimulating Effects of Angelica Gigas by Radio-Protective (당귀의 방사선 방호에 의한 생체면역 증진 효과)

  • Kim Kyung-Yoon;Jeong Hyun-Woo;Kim Gye-Yeop
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.5
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    • pp.1256-1260
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    • 2005
  • The purpose of this study was to investigate the effects of Angelica gigas on jejunal survival, endogenous spleen colony formation and jejunal crypt cells of mice irradiated with Gamma-ray irradiation. The subject of this study includes 42 mice which were divided into each 7 groups. Angelica gigas experiment groups were Angelica gigas + Gamma-ray(10Gy), Angelica gigas + Gamma-ray(3Gy), Angelica gigas. Gamma-ray(1 Gy), Gamma-ray control (10Gy), Gamma-ray control(3Gy), Gamma-ray control(1Gy), Normal groups. In the present study to evaluate the effect of Angelica gigas on jejunal crypt survival, endogenous spleen colony formation, and apoptosis in jejunal crypt cells of mice Gamma-ray with each dose of Gamma-ray irradiation. The results of this study were as follows: In low-dose(1Gy) Gamma-ray radiation were treatment of Angelica gigas showed significantly increased(p<0.05) on the cell death apoptosis in crypt, intestine crypts survival of intestine after gamma-ray irradiation. High-dose(10Gy) Gamma-ray, treatment of Angelica gigas showed significantly increased(p<0.05) on the leukocyte. The above results suggest that Angelica gigas were immunostimulating effectively reduced Gamma-ray irradiation.

Protective effects of Betula platyphylla var. japonica extracts against the cellular damage induced by reactive oxygen species

  • Ji, Sang-Jin;Kim, Jeong-Hee
    • International Journal of Oral Biology
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    • v.34 no.1
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    • pp.15-20
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    • 2009
  • In our present study, total methanol extracts prepared from B. platyphylla var. japonica showed a significant increase in cell proliferation upon the induction of oxidative stress by hydrogen peroxide or $\gamma$-ray irradiation. Total methanol extracts were fractionated into five separate preparations i.e. n-hexane, dichloromethane, ethylacetate, n-butanol and water fractions. Among these, the ethylacetate and butanol fractions of B. platyphylla var. japonica showed the highest protective effects against oxidative stress induced by hydrogen peroxide. These fractions also showed strong protective effects against $\gamma$-ray irradiation. When we evaluated the cytotoxicity of these fractions, the butanol fraction showed no effects in a colony formation assay. In addition, the butanol fraction showed a cell proliferation activation effect evidenced by significant increase in the colony formation of $\gamma$-ray irradiated cells. Both a radical scavenging activity and clonogenic activity assay suggested that the mechanism behind this protective effect against reactive oxygen species may be due to the radical scavenging and cell proliferation activity of B. platyphylla var. japonica extracts.

The Effects of Shi-Quan-Dai-Bu-Tang and Its Ingredients on the Survival of Jejunal Crypt Cells and Hematopoietic Cells in Irradiated Mice (방사선 피폭 마우스에서 소장움세포 및 조혈세포 생존에 미치는 십전대보탕 및 구성단미의 영향)

  • Jo, Sung-Kee;Yu, Young-Bub;Oh, Heon;Kim, Se-Ra;Kim, Sung-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.1
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    • pp.93-98
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    • 2000
  • We performed this study to determine the radioprotective effects of Shi-Quan-Dai-Bu-Tang, as a prescription of traditional Oriental medicine, and its major ingredients. The jejunal crypt survival, endogenous spleen colony formation, and apoptosis in jejunal crypt cells were investigated in mice irradiated with high and low dose of ${\gamma}$-rays. The administration of danggui, baishaoyao, rensan, gancao or baifuling before irradiation protected the jejunal crypts (p<0.005). Shoudehuang, danggui, baishaoyao, rensan and huangqui increased the formation of endogenous spleen colony (p<0.05). Chuanxiong, baishaoyao, rensan and baifuling reduced the frequency of radiation-induced apotosis (p<0.05). The results indicated that the extracts of danggui, baishaoyao, rensan and baifuling may have radoprotective effects in mice irradiated with high and low dose of ${\gamma}$-rays. The radioprotective effect of the prescription, Shi-Qaun-Da-Bu-Tang, was not significant.

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