Journal: Journal of Cancer

Article Title: Inhibition of the cell migration, invasion and chemoresistance of colorectal cancer cells through targeting KLF3 by miR-365a-3p

doi: 10.7150/jca.61967

Figure Lengend Snippet: Expression levels of miR-365a-3p are downregulated in CRC tissues and cell lines. (A) RT-qPCR was used to analyze miR-365a-3p expression levels in 162 CRC and adjacent normal tissues. (B) Log2 transformed value of miR-365a-3p expression level ratio between CRC and adjacent normal tissues. (C) RT-qPCR was used to determine miR-365a-3p expression levels in different CRC cell lines (HCT8, DLD1, LoVo, SW48, HT29, SW480, RKO and HCT116) and a human normal fetal colonic mucosa cell line (FHC). RT-qPCR was performed to analyze miR-365a-3p expression levels in CRC samples with or without (D) lymph node metastasis or (E) distant organ metastasis. (F) Association between relative miR-365a-3p expression levels in CRC tumor tissue and tumor volume of surgically resected clinical samples. RT-qPCR, reverse transcription-quantitative PCR; miR, microRNA; CRC, colorectal cancer. **P<0.05 compared with group FHC, *** P<0.01 compared with group FHC.

Article Snippet: CRC cell lines (HCT8, DLD1, LoVo, SW48, HT29, SW480, RKO and HCT116) and a human normal fetal colonic mucosa cell line (FHC) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Quantitative RT-PCR, Transformation Assay, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Journal of Cancer

Article Title: Inhibition of the cell migration, invasion and chemoresistance of colorectal cancer cells through targeting KLF3 by miR-365a-3p

doi: 10.7150/jca.61967

Figure Lengend Snippet: Effect of miR-365a-3p on the migration and invasion of CRC cell lines. (A) RT-qPCR was used to analyze miR-365a-3p expression levels in SW480 and LOVO cells transfected with three different miR-365a-3p-specific inhibitors or mimics, respectively. (B) RT-qPCR was used to determine miR-365a-3p expression levels in SW480 and LOVO cells transfected with lentiviral vectors carrying a miR-365a-3p-specific inhibitor or mimic, respectively. (C-F) Migratory and invasive abilities of SW480 and LOVO cells transfected with lentiviral vectors carrying a miR-365a-3p-specific inhibitor or mimic, respectively, were determined. RT-qPCR, reverse transcription-quantitative PCR; miR, microRNA. **P<0.05 compared with group INC, mNC, LV-INC, or LV-mNC. *** P<0.01 compared with group INC, mNC, LV-INC, or LV-mNC.

Article Snippet: CRC cell lines (HCT8, DLD1, LoVo, SW48, HT29, SW480, RKO and HCT116) and a human normal fetal colonic mucosa cell line (FHC) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Migration, Quantitative RT-PCR, Expressing, Transfection, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Journal of Cancer

Article Title: Inhibition of the cell migration, invasion and chemoresistance of colorectal cancer cells through targeting KLF3 by miR-365a-3p

doi: 10.7150/jca.61967

Figure Lengend Snippet: Interference of miR-365a-3p expression alters the viability of CRC cells following treatment with chemotherapy agents. Viability of LOVO cells transfected with a lentiviral vector carrying a miR-365a-3p-specific mimic and treated with increasing doses of (A) doxorubicin, (B) fluorouracil or (C) cisplatin. Viability of SW480 cells transfected with a lentiviral vector carrying a miR-365a-3p-specific inhibitor and treated with increasing doses of (D) doxorubicin, (E) fluorouracil or (F) cisplatin. miR, microRNA. **P<0.05 compared with group LV-INC or LV-mNC. *** P<0.01 compared with group LV-INC or LV-mNCFigure 3. Interference of miR-365a-3p expression alters the viability of CRC cells following treatment with chemotherapy agents. Viability of LOVO cells transfected with a lentiviral vector carrying a miR-365a-3p-specific mimic and treated with increasing doses of (A) doxorubicin, (B) fluorouracil or (C) cisplatin. Viability of SW480 cells transfected with a lentiviral vector carrying a miR-365a-3p-specific inhibitor and treated with increasing doses of (D) doxorubicin, (E) fluorouracil or (F) cisplatin. miR, microRNA. **P<0.05 compared with group LV-INC or LV-mNC. *** P<0.01 compared with group LV-INC or LV-mNC.

Article Snippet: CRC cell lines (HCT8, DLD1, LoVo, SW48, HT29, SW480, RKO and HCT116) and a human normal fetal colonic mucosa cell line (FHC) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Transfection, Plasmid Preparation

Journal: Journal of Cancer

Article Title: Inhibition of the cell migration, invasion and chemoresistance of colorectal cancer cells through targeting KLF3 by miR-365a-3p

doi: 10.7150/jca.61967

Figure Lengend Snippet: KLF3 is a target gene of miR-365a-3p and modulates chemoresistance in CRC cell lines. (A) miR-365a-3p binding site in the WT-KLF3 3'-UTR region was predicted and vectors carrying WT- and MUT-KLF3 3'-UTR region were synthesized. (B and C) Dual luciferase reporter gene assay was conducted to investigate the effect on KLF3 expression in LOVO and SW480 cells co-transfected with the lentiviral vectors carrying a miR-365a-3p-specific inhibitor or mimic, respectively, and WT- or MUT-KLF3 3'-UTR vectors. (D) Western blotting and reverse transcription-quantitative PCR were used to determine the regulatory effects of KLF3-specific siRNAs on KLF3 protein and mRNA expression levels in LOVO cells, respectively. (E-G) Viability of SW480 cells transfected with a lentiviral vector carrying a miR-365a-3p-specific inhibitor, with or without the co-transfection with KLF3-specific siRNAs. Each group of cells was treated with increasing doses of fluorouracil, cisplatin or doxorubicin. (H and I) Cell migration and invasion were measured following transfection with a lentiviral vector carrying a miR-365a-3p-specific inhibitor, with or without co-transfection with KLF3-specific siRNAs. miR, microRNA; WT, wild-type; MUT, mutant; KLF3, Kruppel-like factor; UTR, untranslated region; siRNA, small interfering RNA. **P<0.05 compared with group NC, LV-mNC, or LV-INC. *** P<0.01 compared with group NC, LV-mNC, or LV-INC.

Article Snippet: CRC cell lines (HCT8, DLD1, LoVo, SW48, HT29, SW480, RKO and HCT116) and a human normal fetal colonic mucosa cell line (FHC) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Binding Assay, Synthesized, Luciferase, Reporter Gene Assay, Expressing, Transfection, Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction, Plasmid Preparation, Cotransfection, Migration, Mutagenesis, Small Interfering RNA

Journal:

Article Title: Enterotoxigenic Escherichia coli TibA Glycoprotein Adheres to Human Intestine Epithelial Cells

doi: 10.1128/IAI.69.1.52-57.2001

Figure Lengend Snippet: Binding of purified and biotinylated TibA to HCT8 monolayers. Bound TibA was detected by peroxidase-coupled streptavidin. Color development in each well was measured at 405 nm. Symbols: ♦, saturation curve performed with 0.0, 1.0, 2.4, 4.8, 9.6, 19.2, and 48.0 pmol of TibA per well; ▴, competition curve performed with 4.8 pmol of labeled TibA plus 0.0, 1.0, 4.8, 9.6, or 48.0 pmol of unlabeled TibA. Every experiment was performed at least three times. The extent of biotin labeling varied from day to day, resulting in significant differences in color development. Therefore, both curves shown here represent the average of two experiments performed in duplicate.

Article Snippet: The human ileocecal epithelial cell line HCT8 (ATCC CCL 244) was maintained in RPMI 1640 medium containing 10% fetal bovine serum, 1 mM l -glutamine, and 1 mM sodium pyruvate (Life Technologies).

Techniques: Binding Assay, Purification, Labeling

Journal:

Article Title: Enterotoxigenic Escherichia coli TibA Glycoprotein Adheres to Human Intestine Epithelial Cells

doi: 10.1128/IAI.69.1.52-57.2001

Figure Lengend Snippet: Inhibition of TibA-mediated invasion by anti-TibA antiserum. Invasion assays were performed in the absence of antibodies (▪) or in the presence of preabsorbed and affinity-purified IgG from rabbit preimmune serum or polyclonal anti-TibA antiserum. (A) Invasion of HCT8 cells relative to E. coli DH5α(pET109), representing 100% (actual invasion of this strain in the absence of antibody was 1.30% ± 0.05%). Statistically significant effects of antibody treatment on the invasion efficiency of DH5α(pET109) are indicated by ∗ (P < 0.01) or ∗∗ (P < 0.005) as determined by analysis of variance of three experiments, each performed in triplicate. (B) Invasion of HCT8 cells relative to ETEC strain H10407, representing 100% (actual invasion of this strain in the absence of antibody was 0.24% ± 0.02%). TIB3 is a tib locus deletion mutant of H10407. Data are shown as averages for three replicates. Statistically significant effects of antibody treatment on the invasion efficiency of H10407 are indicated by ∗ (P < 0.01) as determined by analysis of variance of three experiments, each performed in triplicate.

Article Snippet: The human ileocecal epithelial cell line HCT8 (ATCC CCL 244) was maintained in RPMI 1640 medium containing 10% fetal bovine serum, 1 mM l -glutamine, and 1 mM sodium pyruvate (Life Technologies).

Techniques: Inhibition, Affinity Purification, Mutagenesis