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: Molecular Medicine Reports

Article Title: Amplified interleukin-15 expression vectors for cancer immunogene therapy

doi: 10.3892/mmr.1.3.369

Figure Lengend Snippet: Figure 2. IL-15 expression by pHi2-IL15-CMV-tat-transfected cells. SW480 and MCF-7 cells were plated at a concentration of 5x105 cells/well in 2 ml of medium on 6-well plates. After 24 h, cells were transfected with IL-15 expression plasmid pHi2-IL15-CMV-tat using Lipofectamine 2000. Culture supernatants were collected at 24 and 48 h after transfection, and IL-15 expression was detected by ELISA. The mean ± SD of three triplicate experiments are shown.

Article Snippet: Human colon carcinoma SW480 cells and human breast carcinoma MCF-7 cells were obtained from American Type Culture Collection (Manassas, VA).

Techniques: Expressing, Transfection, Concentration Assay, Plasmid Preparation, Enzyme-linked Immunosorbent Assay

Journal: Molecular Medicine Reports

Article Title: Amplified interleukin-15 expression vectors for cancer immunogene therapy

doi: 10.3892/mmr.1.3.369

Figure Lengend Snippet: Figure 4. CEA-specific IL-15 expression by pHi2-spIL15-CEA-tat. SW480 and MCF-7 cells were plated at a concentration of 5x105 cells/well in 2 ml of medium on 6-well plates. After 24 h, cells were transfected with pHi2-spIL15- CEA-tat using Lipofectamine 2000. Culture supernatants were collected at 24 and 48 h after transfection, and IL-15 expression was detected by ELISA. The mean ± SD of three triplicate experiments are shown.

Article Snippet: Human colon carcinoma SW480 cells and human breast carcinoma MCF-7 cells were obtained from American Type Culture Collection (Manassas, VA).

Techniques: Expressing, Concentration Assay, Transfection, Enzyme-linked Immunosorbent Assay

Journal: Molecular Medicine Reports

Article Title: Amplified interleukin-15 expression vectors for cancer immunogene therapy

doi: 10.3892/mmr.1.3.369

Figure Lengend Snippet: Figure 5. IL-15 expression of SW480 cells transfected with pHi2-spIL-15- CEA-tat and pHi2-spIL-15-CMV-tat. SW480 cells were plated at a concen- tration of 5x105 cells/well in 2 ml of medium on 6-well plates. After 24 h, cells were separately transfected with pHi2-spIL-15-CEA-tat and pHi2-spIL- 15-CMV-tat using Lipofectamine 2000. Culture supernatants were collected at 24 and 48 h after transfection, and IL-15 expression was detected by ELISA. The mean ± SD of three triplicate experiments are shown.

Article Snippet: Human colon carcinoma SW480 cells and human breast carcinoma MCF-7 cells were obtained from American Type Culture Collection (Manassas, VA).

Techniques: Expressing, Transfection, Enzyme-linked Immunosorbent Assay

Journal: Molecular Medicine Reports

Article Title: Amplified interleukin-15 expression vectors for cancer immunogene therapy

doi: 10.3892/mmr.1.3.369

Figure Lengend Snippet: Figure 3. Increased IL-15 expression by the replacement of the IL-15 with IL-2 signal sequence. SW480 (A) and MCF-7 (B) cells were plated at a concentration of 5x105 cells/well in 2 ml of medium on 6-well plates. After 24 h, cells were separately transfected with the plasmids pHi2-spIL15-CMV- tat and pHi2-IL15-CMV-tat using Lipofectamine 2000. Culture supernatants were collected at 24 and 48 h after transfection, and IL-15 expression was detected by ELISA. The mean ± SD of three triplicate experiments are shown.

Article Snippet: Human colon carcinoma SW480 cells and human breast carcinoma MCF-7 cells were obtained from American Type Culture Collection (Manassas, VA).

Techniques: Expressing, Sequencing, Concentration Assay, Transfection, Enzyme-linked Immunosorbent Assay

Journal: Lab on a chip

Article Title: A microfluidic array with cellular valving for single cell co-culture.

doi: 10.1039/c0lc00172d

Figure Lengend Snippet: Fig. 6 Heterotypic single cell co-culture. Unlabelled single SW480 cells co-cultured with single SW480 cells metabolically labelled with the fluorescent product of calcein AM (A). A single cell couple (B), and the same couple immunostained with connexin 43 (red, C). A trap : cell diameter ratio of $2.0 was required for efficient (70%) cell coupling (D).

Article Snippet: Human SW480 epithelial, HT29 colon carcinoma and MCF-7 epithelial-like breast cancer cells were purchased from DSMZ (Germany).

Techniques: Co-Culture Assay, Cell Culture, Metabolic Labelling