• Title/Summary/Keyword: Transwell

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In Vitro Maturation, Fertilization and Development of Mouse Oocytes Derived from In Vitro Grown Preantyal Follicles (체외성장된 Preantral Follicle에서 유래된 생쥐난자의 체외성숙, 수정 및 발달)

  • Kim, D.H.;Lee, H.J.;K.S. Chung;Lee, H.T.
    • Korean Journal of Animal Reproduction
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    • v.26 no.2
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    • pp.133-142
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    • 2002
  • The objective of this study was to examine maturation, fertilization and developmental rate of the in vitro-grown mouse oocytes, and to compare these results with those of oocytes grown and matured in vivo. The preantral follicles isolated from 12-day-old mice were cultured on Transwell-COL membrane inserts. After in vitro growth and maturation, 72.5 % of oocytes grown in vitro produced polar body which can be comparable to in vivo growth (70.5 %). However, the mean oocyte diameter of the in vitro group (69.6$\pm$2.1$\mu$m) was smaller than that of the in vivo group (73.3$\pm$3.0$\mu$m). The fertilization rate was significantly lower (p<0.05) in the in vitro group (76.5%) than in the in vivo group (90.2%), however, there was no difference in the percentage of monospermic and polyspermic oocytes between two groups. The capacities of in vitro grown ova to cleave and develop to blastocyst were (57.8 and 14.4%, respectively) significantly lower (p<0.001) than those of the in vivo counterpart (84.4 and 56.6%, respectively). Moreover, the mean number of cells per blastocyst was significantly lower (p<0.05) in the in vitro group (39.0$\pm$10.8) than in the in vivo group (60.5$\pm$12.5). Live young were produced from transferred 2-cell embryos derived from in vitro-grown and matured oocytes. In conclusion, the results show that in vitro-grown oocytes did not achieve the developmental capacity of in vitro-grown oocytes.

S100A14 Promotes the Growth and Metastasis of Hepatocellular Carcinoma

  • Zhao, Fu-Tao;Jia, Zhan-Sheng;Yang, Qun;Song, Le;Jiang, Xiao-Jing
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.6
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    • pp.3831-3836
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    • 2013
  • Background: S100A14 has recently been implicated in the progress of several types of cancers. This study aimed to investigate the clinical significance and possible mechanisms of action of S100A14 in the invasion and metastasis of hepatocellular carcinoma (HCC). Methods: S100A14 expression in HCC was detected at mRNA and protein levels and its prognostic significance was assessed. Functional roles of S100A14 in HCC were investigated using MTT, BrdU, wound healing, transwell invasion assay and HCC metastatic mouse model. Results: S100A14 was significantly elevated in HCC tissues, correlated with multiple tumor nodes, high Edmondson-Steiner grade and vascular invasion. Multivariate Cox analysis showed that the S100A14 expression level was a significant and independent prognostic factor for overall survival (OS) of HCC patients (hazard ratio=1.98, 95% confidence interval=1.14-3.46, P=0.013). S100A14 promoted cell proliferation, invasion and metastasis of HCC in vitro and in vivo. Conclusion: These results suggest S100A14 is a novel prognostic marker and therapeutic target for HCC.

ZNF217 is Overexpressed and Enhances Cell Migration and Invasion in Colorectal Carcinoma

  • Zhang, Zi-Chao;Zheng, Li-Qiang;Pan, Li-Jie;Guo, Jin-Xing;Yang, Guo-Shan
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.6
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    • pp.2459-2463
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    • 2015
  • Background: To investigate the expression and clinical significance of zinc finger protein 217 (ZNF217) in human colorectal carcinoma (CRC). Materials and Methods: The expression of ZNF217 in 60 CRC tissues and matched tumor adjacent tissues, collected between January 2013 and June 2014, was assessed immunohistochemically. The relationship between the expression of ZNF217 and clinicopathlogical features was analyzed by Pearson chi-square test. In addition, siRNA was used to down-regulate the expression of ZNF217 in CRC cells. The effects of ZNF217 for cell migration and invasion were measured by wound healing assay and transwell assay, respectively. Results: The expression level of ZNF217 was significantly higher in CRC tissues than in tumor adjacent tissues (p<0.05), positively correlating with tumor size, lymphatic metastasis and advanced TNM stage (p<0.05). Down-regulation of ZNF217 in CRC cells could significantly suppress cell migration and invasion. Conclusions: ZNF217 is overexpressed in colorectal carcinoma tissues and is associated with tumor malignant clinicopathological features. ZNF217 may promote CRC progression by inducing cell migration and invasion.

Inhibition of Metastasis and Invasion of Ovarian Cancer Cells by Crude Polysaccharides from Rosa Roxburghii Tratt in Vitro

  • Chen, Yang;Liu, Zhong-Jing;Liu, Jia;Liu, Li-Kun;Zhang, Er-Shao;Li, Wei-Ling
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10351-10354
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    • 2015
  • Background: Rosa Roxburghii Tratt is a promising wild fruit crop in Southwest China. Its extracts have been used as traditional Chinese medicine, which benefit immune responses and cure various health disorders. However, whether Rosa Roxburghii Tratt polysaccharides could inhibit metastasis and invasion of ovarian cancer cells remains unknown. Materials and Methods: Effects of crude polysaccharides from Rosa Roxburghii Tratt on the viability of ovarian cancer A2780 cells were detected by MTT assay. Ovarian carcinoma cell migration and invasion after exposure to Rosa Roxburghii Tratt polysaccharides were quantified by wound healing and Transwell assays, respectively. Western blotting was applied to assess protein levels of MMP-9. Results: The results indicated that Rosa Roxburghii Tratt polysaccharides significantly reduced wound closure rate of A2780 cells, inhibited their migration and invasion, and suppressed the expression of MMP-9. Conclusions: Our findings indicated that Rosa Roxburghii Tratt polysaccharides have potential for develop as anti-metastatic cancer drug preparations for ovarian cancer patients.

Inhibition of SMP30 Gene Expression Influences the Biological Characteristics of Human Hep G2 Cells

  • Zhang, Sheng-Chang;Liang, Ming-Kang;Huang, Guang-Lin;Jiang, Kui;Zhou, Su-Fang;Zhao, Shuang
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.3
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    • pp.1193-1196
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    • 2014
  • Senescence marker protein 30 (SMP30), a hepatocellular carcinoma (HCe) associated antigen had been identified by our research group. To study its mechanisms of regulation and associations with the occurrence and development of HCe, we inhibited expression by RNAi technique, and observed effects on the biological characteristics of Hep G2 cells. In cell viability assays, cell growth in the experimental group (with siRNA transfection) was elevated. In Transwell invasion assays, compared with blank and control groups, numbers of invading cells in the experimental group were significantly increased, whereas in apoptosis assays, the percentage apoptosis demonstrated no differences, but after UV irradiation, that in the experimental group was higher than the other two groups. In a word, SMP30 can inhibit the proliferation and invasion of human hepatoma cells and thus can be regarded as a cancer suppressive factor.

Increased Migration and Local Invasion Potential of SiHa Cervical Cancer Cells Expressing Aquaporin 8

  • Shi, Yong-Hua;Rehemu, Nijiati;Ma, Hong;Tuokan, Talafu;Chen, Rui;Suzuke, Lalai
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.3
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    • pp.1825-1828
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    • 2013
  • Overexpression of several aquaporins (AQPS) has been reported in different types of human cancer but roles in human carcinogenesis have yet to be clearly defined. Here, we up-regulated expression of the AQP8 gene in SiHa human cervical cancer cells with a lentivirus transfection system and investigated its effects as a potential therapeutic target for cervical cancer. Results showed AQP8 overexpression did not affect their substrate adherence and proliferation, but accelerated migration as assessed by transwell migration and wound healing assays. Moreover, AQP8 overexpression significantly enhanced local invasion of SiHa cells in nude mice. These findings altogether indicate that AQP8 overexpression increases migration of SiHa cells and probably participates in the process of tumor local invasion.

Type 3 muscarinic acetylcholine receptor stimulation is a determinant of endothelial barrier function and adherens junctions integrity: role of protein-tyrosine phosphatase 1B

  • Jiao, Zhou-Yang;Wu, Jing;Liu, Chao;Wen, Bing;Zhao, Wen-Zeng;Du, Xin-Ling
    • BMB Reports
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    • v.47 no.10
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    • pp.552-557
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    • 2014
  • The main purpose of this study was to investigate whether type 3 muscarinic acetylcholine receptor (M3R) dysfunction induced vascular hyperpermeability. Transwell system analysis showed that M3R inhibition by selective antagonist 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP) and small interfering RNA both increased endothelial permeability. Using coimmunoprecipitation and Western blot assay, we found that M3R inhibition increased VE-cadherin and ${\beta}$-catenin tyrosine phosphorylation without affecting their expression. Using PTP1B siRNA, we found that PTP1B was required for maintaining VE-cadherin and ${\beta}$-catenin protein dephosphorylation. In addition, 4-DAMP suppressed PTP1B activity by reducing cyclic adenosine monophosphate (cAMP), but not protein kinase $C{\alpha}$ ($PKC{\alpha}$). These data indicate that M3R preserves the endothelial barrier function through a mechanism potentially maintaining PTP1B activity, keeping the adherens junction proteins (AJPs) dephosphorylation.

Pristimerin Inhibits Breast Cancer Cell Migration by Up-regulating Regulator of G Protein Signaling 4 Expression

  • Mu, Xian-Min;Shi, Wei;Sun, Li-Xin;Li, Han;Wang, Yu-Rong;Jiang, Zhen-Zhou;Zhang, Lu-Yong
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.4
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    • pp.1097-1104
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    • 2012
  • Background/Aim: Pristimerin isolated from Celastrus and Maytenus spp can inhibit proteasome activity. However, whether pristimerin can modulate cancer metastasis is unknown. Methods: The impacts of pristimerin on the purified and intracellular chymotrypsin proteasomal activity, the levels of regulator of G protein signaling 4 (RGS 4) expression and breast cancer cell lamellipodia formation, and the migration and invasion were determined by enzymatic, Western blot, immunofluorescent, and transwell assays, respectively. Results: We found that pristimerin inhibited human chymotrypsin proteasomal activity in MDA-MB-231 cells in a dose-dependent manner. Pristimerin also inhibited breast cancer cell lamellipodia formation, migration, and invasion in vitro by up-regulating RGS4 expression. Thus, knockdown of RGS4 attenuated pristimerin-mediated inhibition of breast cancer cell migration and invasion. Furthermore, pristimerin inhibited growth and invasion of implanted breast tumors in mice. Conclusion: Pristmerin inhibits proteasomal activity and increases the levels of RGS4, inhibiting the migration and invasion of breast cancer cells.

Synergistic Anti-tumor Effect of KLF4 and Curcumin in Human Gastric Carcinoma Cell Line

  • Ji, Jun;Wang, He-Shuang;Gao, Yan-Yan;Sang, Li-Min;Zhang, Li
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.18
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    • pp.7747-7752
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    • 2014
  • Kr$\ddot{u}$ppel-like factor 4 is a transcription factor which plays an important role in development and progression of various carcinomas. Curcumin characterized by excellent anti-cancer properties is regarded as a serviceable natural compound used in carcinoma therapy. This study aimed at exploring the impact of KLF4 overexpression in cooperation with curcumin on the proliferation, apoptosis and invasion of human gastric carcinoma BGC-823 cells. Flow cytometry analysis, CCK-8 assays, transwell assays and Western blot results showed that KLF4 overexpression combined with curcumin had significant anti-proliferation, pro-apoptosis and anti-invasion effects on BGC-823 cells. We also found that KLF4 had synergistic effects with curcumin, better promoting apoptosis and inhibiting proliferation and invasion of gastric carcinona cells. These results indicate that KLF4 could be used as a potential therapeutic target; curcumin could act as an auxiliary and provide a promising therapeutic strategy in stomach cancer.

MiR-150-5p Suppresses Colorectal Cancer Cell Migration and Invasion through Targeting MUC4

  • Wang, Wei-Hua;Chen, Jie;Zhao, Feng;Zhang, Bu-Rong;Yu, Hong-Sheng;Jin, Hai-Ying;Dai, Jin-Hua
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.15
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    • pp.6269-6273
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    • 2014
  • Growing evidence suggests that miR-150-5p has an important role in regulating genesis of various types of cancer. However, the roles and the underlying mechanisms of miR-150-5p in development of colorectal cancer (CRC) remain largely unknown. Transwell chambers were used to analyze effects on cell migration and invasion by miR-150-5p. Quantitative real-time PCR (qRT-PCR), Western blotting and dual-luciferase 3' UTR reporter assay were carried out to identify the target genes of miR-150-5p. In our research, miR-150-5p suppressed CRC cell migration and invasion, and MUC4 was identified as a direct target gene. Its effects were partly blocked by re-expression of MUC4. In conclusiomn, miR-150-5p may suppress CRC metastasis through directly targeting MUC4, highlighting its potential as a novel agent for the treatment of CRC metastasis.