Journal: International Journal of Molecular Sciences

Article Title: Epigenetic Silencing of miR-218-5p Modulates BIRC5 and DDX21 Expression to Promote Colorectal Cancer Progression

doi: 10.3390/ijms26094146

Figure Lengend Snippet: Validation of selected miR-218-5p Gene Targets. ( A ) Validation of BIRC5 and DDX21 as bona fide gene targets for miR-218-5p in CRC. Two-tailed t -test was used to compare different groups. ** p < 0.005, *** p < 0.0005, **** p < 0.00005. ( B ) Western blot showing DDX21 protein expression in miR-218-5p overexpressing compared to control HT-29 and HCT116 cells. Quantification of DDX21 protein expression normalized to ACTB is shown in the right panel. ( C ) Gene effect score based on CRISPR-Cas9 screen data in 40 CRC cell models from the DepMap database.

Article Snippet: A notable observation within SLIT2 , which encodes miR-218-1 within its intronic region, is the detection of higher levels of hypermethylation near the end of the promoter regions in the CRC cell lines, just upstream of the transcription start site, where the TATA box is typically located.

Techniques: Biomarker Discovery, Two Tailed Test, Western Blot, Expressing, Control, CRISPR

Journal: International Journal of Molecular Sciences

Article Title: Epigenetic Silencing of miR-218-5p Modulates BIRC5 and DDX21 Expression to Promote Colorectal Cancer Progression

doi: 10.3390/ijms26094146

Figure Lengend Snippet: Suppression of SLIT2 and SLIT3 miR-218 Host Genes in CRC. ( A ) Genomic location of miR-218-1 and miR-218-2 within the SLIT2 and SLIT3 genomic region on chromosome 4 and 5, respectively. ( B ) Correlation plot between miR-218-5p and SLIT2 ( left ) and SLIT3 ( right ) in a large cohort of COAD (n = 450) from the ENCORI project. ( C ) Downregulation of SLIT2 ( left ) and SLIT3 ( right ) in COAD (n = 275) compared to normal colon tissue (n = 349) from GEPIA2 database. T: tumor, N: normal. * p < 0.05. ( D ) Methylation analysis of SLIT2 and SLIT3 promoters using bisulfite conversion and NGS in a panel of CRC cell models (HCT116, HT-29, SW-480, LoVo, and DLD-1) compared to the MCF10A normal epithelial cells. ( E ) Schematic representation illustrating (🠯) downregulation of SLIT2 and SLIT3 in CRC to lead to miR-218-5p suppression (🠯), thus promoting tumorigenesis due to lifted suppression (🠭) of BIRC5, DDX21, and other gene targets identified in the current study.

Article Snippet: A notable observation within SLIT2 , which encodes miR-218-1 within its intronic region, is the detection of higher levels of hypermethylation near the end of the promoter regions in the CRC cell lines, just upstream of the transcription start site, where the TATA box is typically located.

Techniques: Methylation

Journal: Journal of Translational Medicine

Article Title: Disruption of the c-Myc/miR-200b-3p/PRDX2 regulatory loop enhances tumor metastasis and chemotherapeutic resistance in colorectal cancer

doi: 10.1186/s12967-017-1357-7

Figure Lengend Snippet: MiR-200b-3p inhibits CRC cells’ metastatic capacity by targeting PRDX2 in vivo. a Intestinal and hepatic metastatic nodules after subcutaneous tumors derived from LoVo/NC, LoVo/miR and LoVo/miR + PRDX2 cells were transplanted in the mesentery at the distal end of cecum in mice (n = 5) for 6 weeks. Red arrows point at potential metastatic nodules in intestines. Scale bars represent 50 μm. b , c The number of hepatic metastatic nodules ( b ) or intestinal metastatic nodules ( c ) of mice with tumors derived from LoVo/NC, LoVo/miR and LoVo/miR + PRDX2 cells. The number of hepatic metastatic nodules per mouse was counted under the microscope, with five high power fields (HPF) observation (*** p < 0.001). d Intestinal and hepatic metastatic nodules after subcutaneous tumors derived from SW480/NC and SW480/Zip-miR cells were transplanted in the mesentery at the distal end of cecum in mice (n = 5) for 6 weeks. Red arrows point at potential metastatic nodules in intestines. Scale bars represent 50 μm. e , f The number of hepatic metastatic nodules ( e ) or intestinal metastatic nodules ( f ) of mice with tumors derived from SW480/NC and SW480/Zip-miR cells. The number of hepatic metastatic nodules per mouse was counted under the microscope, with five HPF observation (*** p < 0.001)

Article Snippet: First, 1 × 10 6 CRC cells with different treatments were subcutaneously injected into the female BALB/c nude mice (Beijing HFK Bioscience Co., LTD, Beijing China) aged 5 weeks for 4 weeks to generate subcutaneous tumors and then the mice were sacrificed.

Techniques: In Vivo, Derivative Assay, Microscopy

Journal: Journal of Translational Medicine

Article Title: Disruption of the c-Myc/miR-200b-3p/PRDX2 regulatory loop enhances tumor metastasis and chemotherapeutic resistance in colorectal cancer

doi: 10.1186/s12967-017-1357-7

Figure Lengend Snippet: Disruption of the c-Myc/miR-200b-3p/PRDX2 regulatory loop enhances chemotherapeutic resistance of CRC cells. a c-Myc overexpression enhanced the resistance of SW480 cells to oxaliplatin, while the enhanced effect was dampened by miR-200b-3p partly. b miR-200b-3p overexpression reduced the resistance of LoVo cells to oxaliplatin, while the reduced effect was reversed by nontargetable PRDX2 partly. c c-Myc overexpression repressed oxaliplatin-induced apoptosis in SW480 cells, while the repressive effect was reversed by miR-200b-3p partly. d miR-200b-3p overexpression enhanced oxaliplatin-induced apoptosis in LoVo cells, while the enhanced effect was abolished by nontargetale PRDX2 partly. e , f The oxaliplatin-induced apoptosis was statistically analyzed in LoVo cells ( e ) or in SW480 cells ( f ) among different treatment groups

Article Snippet: First, 1 × 10 6 CRC cells with different treatments were subcutaneously injected into the female BALB/c nude mice (Beijing HFK Bioscience Co., LTD, Beijing China) aged 5 weeks for 4 weeks to generate subcutaneous tumors and then the mice were sacrificed.

Techniques: Disruption, Over Expression

Journal: Oncogenesis

Article Title: Combination curcumin and (−)-epigallocatechin-3-gallate inhibits colorectal carcinoma microenvironment-induced angiogenesis by JAK/STAT3/IL-8 pathway

doi: 10.1038/oncsis.2017.84

Figure Lengend Snippet: Colorectal carcinoma CM enhanced the migration, invasion, tube formation and Dil-Ac-LDL uptake abilities of NECs. ( a ) NECs monolayer was wounded and induced by SW620, HT-29 or HCT116 CM. Photographs were taken after induction for 0, 24 and 48 h (scale bar 40 μm). ( b ) NECs were induced by SW620, HT-29 or HCT116 CM for 48 h. The number of invaded cells was counted in three random fields. Representative images of invaded cells were shown (scale bar 40 μm). ( c ) Formation tubes in each group were photographed. The number of tubes per field was counted in three random fields (scale bar 20 μm). ( d ) Representative images showed the Dil-Ac-LDL uptake ability of NECs and quantification of the relative Dil-Ac-LDL uptake (scale bar 40 μm). Data are presented as mean±s.d. from three independent experiments. *** P <0.001.

Article Snippet: The human colorectal cancer cell lines SW620, HT-29 and HCT116 were obtained from Nanjing Keygen Biotech Corp. (Nanjing, China), and cultured in RPMI-1640 (Biological Industries, Kibbutz Beit Haemek, Israel) medium with 10% fetal bovine serum.

Techniques: Migration

Journal: Oncogenesis

Article Title: Combination curcumin and (−)-epigallocatechin-3-gallate inhibits colorectal carcinoma microenvironment-induced angiogenesis by JAK/STAT3/IL-8 pathway

doi: 10.1038/oncsis.2017.84

Figure Lengend Snippet: Colorectal carcinoma CM promoted the transition of NECs toward TECs. ( a ) Immunohistochemical staining for TECs markers (TEM1, TEM8 and VEGFR2) in colorectal carcinoma and peri-carcinoma tissue (scale bar 20 μm). ( b ) NECs were induced by SW620, HT-29 or HCT116 CM for 48 h. The relative mRNA levels of TECs markers were determined by qRT–PCR. ( c , d ) NECs were induced by SW620, HT-29 or HCT116 CM for 48 h, the protein levels of TECs markers were detected by western blot ( c ) and immunofluorescence ( d ). Data are presented as mean±s.d. from three independent experiments. ** P <0.01, *** P <0.001.

Article Snippet: The human colorectal cancer cell lines SW620, HT-29 and HCT116 were obtained from Nanjing Keygen Biotech Corp. (Nanjing, China), and cultured in RPMI-1640 (Biological Industries, Kibbutz Beit Haemek, Israel) medium with 10% fetal bovine serum.

Techniques: Immunohistochemical staining, Staining, Quantitative RT-PCR, Western Blot, Immunofluorescence

Journal: Oncogenesis

Article Title: Combination curcumin and (−)-epigallocatechin-3-gallate inhibits colorectal carcinoma microenvironment-induced angiogenesis by JAK/STAT3/IL-8 pathway

doi: 10.1038/oncsis.2017.84

Figure Lengend Snippet: JAK/STAT3 signaling pathway was activated during the transition of NECs toward TECs induced by colorectal carcinoma CM. ( a ) NECs were induced by SW620, HT-29 or HCT116 CM for 48 h. The relative mRNA levels of JAK and STAT3 were measured by qRT–PCR. ( b , c ) The expression level of indicated protein was detected by western blot ( b ) and immunofluoresence (scale bar 50 μm) ( c ). Data are expressed as mean±s.d. from three independent experiments. * P <0.05, ** P <0.01, *** P <0.001.

Article Snippet: The human colorectal cancer cell lines SW620, HT-29 and HCT116 were obtained from Nanjing Keygen Biotech Corp. (Nanjing, China), and cultured in RPMI-1640 (Biological Industries, Kibbutz Beit Haemek, Israel) medium with 10% fetal bovine serum.

Techniques: Quantitative RT-PCR, Expressing, Western Blot

Journal: Neoplasia (New York, N.Y.)

Article Title: Biologic Response of Colorectal Cancer Xenograft Tumors to Sequential Treatment with Panitumumab and Bevacizumab

doi: 10.1016/j.neo.2018.04.006

Figure Lengend Snippet: Antitumor effect in LIM1215(A) xenografts by treatment received. (A and B) assessed by average tumor volume and (C) relative growth rate. Arrowheads indicate dosing. Arrows indicate sample harvesting. Data represent mean ± SE. ( n = 3–7. * P < .05). B, bevacizumab; P, panitumumab; V, vehicle control.

Article Snippet: The human colon cancer cell line LIM1215 was obtained from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK).

Techniques: Control

Journal: Neoplasia (New York, N.Y.)

Article Title: Biologic Response of Colorectal Cancer Xenograft Tumors to Sequential Treatment with Panitumumab and Bevacizumab

doi: 10.1016/j.neo.2018.04.006

Figure Lengend Snippet: Outcome of immunohistochemistry for Ki-67 in LIM1215(B) xenograft sections. (A) Using vehicle control, (B) panitumumab-bevacizumab, (C) bevacizumab-panitumumab, and (D) bevacizumab-bevacizumab. (E) Proportion of Ki-67-positive cells in all treatment groups. Sections were IHC stained for Ki-67 (brown) and counterstained with hematoxylin (purple). Representative images of the sections are shown. Data in the graph represent the mean ± SE ( n = 6–8). ** P < .01. B, bevacizumab; P, panitumumab; V, vehicle control.

Article Snippet: The human colon cancer cell line LIM1215 was obtained from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK).

Techniques: Immunohistochemistry, Control, Staining

Journal: Neoplasia (New York, N.Y.)

Article Title: Biologic Response of Colorectal Cancer Xenograft Tumors to Sequential Treatment with Panitumumab and Bevacizumab

doi: 10.1016/j.neo.2018.04.006

Figure Lengend Snippet: Levels of Phosphorylated Growth Factor Receptors in LIM1215(A) Xenografts Treated with PB, BP, and BB Relative to Vehicle Control

Article Snippet: The human colon cancer cell line LIM1215 was obtained from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK).

Techniques: Phospho-proteomics, Control

Journal: Neoplasia (New York, N.Y.)

Article Title: Biologic Response of Colorectal Cancer Xenograft Tumors to Sequential Treatment with Panitumumab and Bevacizumab

doi: 10.1016/j.neo.2018.04.006

Figure Lengend Snippet: Results of western blotting in LIM1215(B) xenografts. (A) EPHA2 and pEPHA2 with panitumumab-bevacizumab compared with bevacizumab-panitumumab. (B) EPHA2 and pEPHA2 with panitumumab-bevacizumab compared with bevacizumab-bevacizumab. (C) Phosphorylation of EPHA2 for panitumumab-bevacizumab compared with bevacizumab-panitumumab and (D) compared with bevacizumab-bevacizumab. (E) RSK and pRSK with panitumumab-bevacizumab compared with bevacizumab-panitumumab and (F) RSK and pRSK with panitumumab-bevacizumab compared with bevacizumab-bevacizumab. (G) Phosphorylation of RSK for panitumumab-bevacizumab compared with bevacizumab-panitumumab and (H) compared with bevacizumab-bevacizumab. Data represent mean ± SD ( n = 8). ** P < .01, *** P < .001. B, bevacizumab; P, panitumumab; V, vehicle control.

Article Snippet: The human colon cancer cell line LIM1215 was obtained from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK).

Techniques: Western Blot, Phospho-proteomics, Control

Journal: Neoplasia (New York, N.Y.)

Article Title: Biologic Response of Colorectal Cancer Xenograft Tumors to Sequential Treatment with Panitumumab and Bevacizumab

doi: 10.1016/j.neo.2018.04.006

Figure Lengend Snippet: Enrichment Analysis of LIM1215(A) Xenografts Treated with Bevacizumab-Bevacizumab Compared with Vehicle Control (all Canonical Pathways, P < .001)

Article Snippet: The human colon cancer cell line LIM1215 was obtained from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK).

Techniques: Control, Activation Assay, Inhibition

Journal: Neoplasia (New York, N.Y.)

Article Title: Biologic Response of Colorectal Cancer Xenograft Tumors to Sequential Treatment with Panitumumab and Bevacizumab

doi: 10.1016/j.neo.2018.04.006

Figure Lengend Snippet: Relative expression of (A–H) lipogenic ( FASN, HMGCR, MVD, LSS ) and (I–L) hypoxia-related ( CA9, TGFBI ) genes in LIM1215(B) xenograft tumors. Expression relative to vehicle control with first-line treatment is shown in (A–D) and (I–J), and with sequential treatment in (E–H) and (K–L). Data represent mean ± SD ( n = 8). * P < .05, ** P < .01. B, bevacizumab; P, panitumumab; V, vehicle control.

Article Snippet: The human colon cancer cell line LIM1215 was obtained from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK).

Techniques: Expressing, Control