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: Theranostics

Article Title: Tumor-intrinsic CD47 signal regulates glycolysis and promotes colorectal cancer cell growth and metastasis

doi: 10.7150/thno.40860

Figure Lengend Snippet: CD47 enhances CRC cell proliferation and metastasis in vivo . (A-H). CD47 promotes growth of CRC cells in vivo. DLD1-Vector, DLD1-CD47, SW480-shScramble, and SW480-shCD47 CRC cells were subcutaneously injected into nude mice (n = 5 and 4 respectively). Images of tumors (A, D), statistics of tumor weights (B, E), tumor volumes (C, F), and representative IHC staining are shown (G, H). (I-N). CD47 increases tumor metastasis in vivo . DLD1-Vector, DLD1-CD47, SW480-shScramble, and SW480-shCD47 cells were injected into tail veins of nude mice (n = 5). Representative images (I, L), HE staining (J, M), and statistics of lung metastatic tumors are depicted (K, N). Data are the means ± SD, * p <0.05, ** p <0.01, *** p <0.001.

Article Snippet: The human embryonic kidney epithelial cell line 293T (HEK293T) and human CRC cell lines (HCT8, DLD1, HCT116, SW480, HT29, HCT15, KM12, SW620, RKO) used were obtained from American type culture collection (ATCC).

Techniques: In Vivo, Plasmid Preparation, Injection, Immunohistochemistry, Staining

Journal: Theranostics

Article Title: Tumor-intrinsic CD47 signal regulates glycolysis and promotes colorectal cancer cell growth and metastasis

doi: 10.7150/thno.40860

Figure Lengend Snippet: Rescue assays of CRC cell proliferation, migration, and invasion in vitro . (A-D). Transwell migration /invasion assays and cell proliferation assays were performed after ENO1 silencing in control and CD47-overexpressed DLD1/HCT8 cells. (E-G). Transwell migration/invasion assays and cell proliferation assays were performed after transiently expressing ENO1 in control and CD47-knockdown HCT116/SW480 cells. Data are means ± SD, * p <0.05, ** p <0.01, *** p <0.001.

Article Snippet: The human embryonic kidney epithelial cell line 293T (HEK293T) and human CRC cell lines (HCT8, DLD1, HCT116, SW480, HT29, HCT15, KM12, SW620, RKO) used were obtained from American type culture collection (ATCC).

Techniques: Migration, In Vitro, Control, Expressing, Knockdown

Journal: Theranostics

Article Title: Tumor-intrinsic CD47 signal regulates glycolysis and promotes colorectal cancer cell growth and metastasis

doi: 10.7150/thno.40860

Figure Lengend Snippet: High expression of CD47 and ENO1 predict poor survival outcomes of CRC patients. (A). Statistical analysis of CD47 mRNA levels of 44 paired samples of CRC and adjacent normal tissues from the Sixth Affiliated Hospital (n = 44, non-parametric Wilcoxon matched-pairs signed rank test, p <0.01). (B). Scatter plot illustrating the protein level (IHC score) of CD47 in CRC and matched adjacent normal tissues. (n=179, p <0.05, Paired Student's t-test). (C-D). Survival analysis of CRC patients layered by the expression of CD47 in tissue microarrays from the Sixth Affiliated Hospital of Sun Yat-sen University (n = 293, Log-rank test, p <0.001, CD47-Low is defined as IHC score < 4 andCD47-High is defined as IHC score >= 4). (E). Scatter plot illustrating the protein level (IHC score) of ENO1 in CRC and matched adjacent normal tissues (n=179, p <0.01, Paired Student's t-test). (F-G). Survival analysis of CRC patients layered by the expression of ENO1 in tissue microarrays from the Sixth Affiliated Hospital of Sun Yat-sen University (n=293, p <0.001, ENO1-Low is defined as IHC score < 5 and ENO1-High is defined as IHC score >= 5). (H). Correlation analysis between CD47 and ENO1 protein levels (IHC score) in CRC samples from the Sixth Affiliated Hospital of Sun Yat-sen University. (n=293, Pearson's correlation, p <0.001). (I). Representative IHC staining of CD47 and ENO1 in human CRC tissues (right) and adjacent normal tissues (left). (J). Survival analysis of CRC patients layered by the expression of CD47 and ENO1 in tissue microarrays from the Sixth Affiliated Hospital of Sun Yat-sen University (n = 293, Log-rank test). (K-L). Receiver operating characteristic (ROC) curve and area under curve (AUC) values for the prognostic prediction model based on CD47 and ENO1 expression. (M). A schematic model of CD47 function in CRC. CD47 interacts with ENO1 and inhibits the FBXW7-mediated degradation of ENO1, thereby increasing ERK phosphorylation and glycolysis, ultimately promoting proliferation and metastasis of CRC.

Article Snippet: The human embryonic kidney epithelial cell line 293T (HEK293T) and human CRC cell lines (HCT8, DLD1, HCT116, SW480, HT29, HCT15, KM12, SW620, RKO) used were obtained from American type culture collection (ATCC).

Techniques: Expressing, Immunohistochemistry, Phospho-proteomics