Journal: Molecular Vision

Article Title: Protein tyrosine phosphatase-1B (PTP1B) helps regulate EGF-induced stimulation of S-phase entry in human corneal endothelial cells

doi:

Figure Lengend Snippet: Western blot detection of PTP1B protein bands in HCEC. Confluent Passage-2 HCEC were cultured from a newborn (lane 1) and a 74-year-old donor (lane 2) and processed for western blot detection of PTP1B. A commercially prepared whole cell lysate of SW480 cells was used as a positive control (lane 3). Blots showed that the two HCEC samples yielded a band at approximately 50 kDa, indicating the presence of full-length PTP1B, as well as two additional bands of approximately 48 kDa and 46 kDa, possibly representing truncated forms. The same three bands were observed in the SW480 positive control sample, as well as lower molecular weight bands around 39–37 kDa that may result from non-specific proteolytic cleavage. Note the variability in density of the three PTP1B-positive bands within the two HCEC samples.

Article Snippet: A commercially prepared whole cell lysate from SW480 cells, a human colonic adenocarcinoma cell line (Santa Cruz Biotechnology Inc., Santa Cruz, CA), acted as a positive control for PTP1B.

Techniques: Western Blot, Cell Culture, Positive Control, Molecular Weight

Journal: Molecular Vision

Article Title: Protein tyrosine phosphatase-1B (PTP1B) helps regulate EGF-induced stimulation of S-phase entry in human corneal endothelial cells

doi:

Figure Lengend Snippet: Comparison of EGFR and PTP1B protein expression in HCEC cultured from 4 young and 4 older donors. A : Western blots demonstrate the relative expression of EGFR and PTP1B in each of the 8 donor samples. EGFR was expressed in all 8 samples, although the band density varied somewhat among the samples. Three PTP1B bands were observed in most samples. These bands are indicated within brackets. Blots for the positive controls for EGFR (EGF-treated A431cells) and PTP1B (SW480 cells) are not shown, but bands were obtained at the same relative molecular weights as in the HCEC samples. β-Actin was used as a loading control for both EGFR and PTP1B. B : Comparison of the average band density for EGFR and PTP1B in samples from young and older donors. Bars represent SEM. Asterisk indicates statistical significance (p=0.024).

Article Snippet: A commercially prepared whole cell lysate from SW480 cells, a human colonic adenocarcinoma cell line (Santa Cruz Biotechnology Inc., Santa Cruz, CA), acted as a positive control for PTP1B.

Techniques: Comparison, Expressing, Cell Culture, Western Blot, Control

Journal: Aging (Albany NY)

Article Title: ESM1 promotes angiogenesis in colorectal cancer by activating PI3K/Akt/mTOR pathway, thus accelerating tumor progression

doi: 10.18632/aging.204559

Figure Lengend Snippet: Detection of migration and invasion abilities in SW480 and SW620 cells. ( A ) Distances of migration at 0, 24 and 48 h after scratch formation in SW480 cells. ( B ) Distances of migration at 0, 24 and 48 h after scratch formation in SW620 cells. ( C ) Number of migrating and invading SW480 cells. ( D ) Number of migrating and invading SW620 cells. * p < 0.05 and ** p < 0.01 vs. ESM1-NC group.

Article Snippet: CRC SW480 and SW620 cell lines and human normal colorectal cells (CCD-841CoN, IM-H477) were purchased from Nanjing KeyGen Biotech Co., Ltd. SW480 and SW620 cells were inoculated into L15 medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin and incubated in an incubator at 37°C with 5% CO 2 .

Techniques: Migration

Journal: Aging (Albany NY)

Article Title: ESM1 promotes angiogenesis in colorectal cancer by activating PI3K/Akt/mTOR pathway, thus accelerating tumor progression

doi: 10.18632/aging.204559

Figure Lengend Snippet: Proliferation of SW480 and SW620 cells in each group. ( A ) Number of colonies formed in SW480 cells transfected with ESM1-NC and ESM1-mimic. ( B ) Number of colonies formed in SW620 cells transfected with ESM1-NC and ESM1-mimic. ( C ) Detection of relative viability of SW480 cells via MTT assay. ( D ) Detection of relative viability of SW620 cells via MTT assay. * p < 0.05 and ** p < 0.01 vs. ESM1-NC group.

Article Snippet: CRC SW480 and SW620 cell lines and human normal colorectal cells (CCD-841CoN, IM-H477) were purchased from Nanjing KeyGen Biotech Co., Ltd. SW480 and SW620 cells were inoculated into L15 medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin and incubated in an incubator at 37°C with 5% CO 2 .

Techniques: Transfection, MTT Assay

Journal: Aging (Albany NY)

Article Title: ESM1 promotes angiogenesis in colorectal cancer by activating PI3K/Akt/mTOR pathway, thus accelerating tumor progression

doi: 10.18632/aging.204559

Figure Lengend Snippet: Relative protein expressions of Cyclin D1 and Cyclin A2 in SW480 and SW620 cells. ( A, B ) Relative protein expressions of Cyclin D1 and Cyclin A2 in ESM1-NC group, ESM1-mimic group, ESM1-NC + PI3K inhibitor group and ESM1-mimic + PI3K inhibitor group. ( C, D ) Statistics for Cyclin D1 and Cyclin A2 protein expressions. * p < 0.05 and ** p < 0.01 vs. ESM1-NC group.

Article Snippet: CRC SW480 and SW620 cell lines and human normal colorectal cells (CCD-841CoN, IM-H477) were purchased from Nanjing KeyGen Biotech Co., Ltd. SW480 and SW620 cells were inoculated into L15 medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin and incubated in an incubator at 37°C with 5% CO 2 .

Techniques:

Journal: Aging (Albany NY)

Article Title: ESM1 promotes angiogenesis in colorectal cancer by activating PI3K/Akt/mTOR pathway, thus accelerating tumor progression

doi: 10.18632/aging.204559

Figure Lengend Snippet: Endothelial angiogenesis ability under SW480 and SW620 stimulation, and detection of effect of ESM1 on the proliferation of SW480 and SW620 cells via flow cytometry. ( A ) Number of endothelial cell tube formation in 3B-11 cells co-cultured with SW480 cells transfected with ESM1-NC and ESM1-mimic. ( B ) Number of endothelial cell tube formation in 3B-11 cells co-cultured with SW620 cells transfected with ESM1-NC and ESM1-mimic. ( C , D ) Flow cytometry reveals that ESM1-mimic can significantly increase the ratio of G2 phase cells and promote cell proliferation. * p < 0.05 and ** p < 0.01 vs. ESM1-NC group.

Article Snippet: CRC SW480 and SW620 cell lines and human normal colorectal cells (CCD-841CoN, IM-H477) were purchased from Nanjing KeyGen Biotech Co., Ltd. SW480 and SW620 cells were inoculated into L15 medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin and incubated in an incubator at 37°C with 5% CO 2 .

Techniques: Flow Cytometry, Cell Culture, Transfection

Journal: Aging (Albany NY)

Article Title: ESM1 promotes angiogenesis in colorectal cancer by activating PI3K/Akt/mTOR pathway, thus accelerating tumor progression

doi: 10.18632/aging.204559

Figure Lengend Snippet: ESM1 promotes the occurrence and development of tumors in nude mice. ( A ) SW480 cells transfected with ESM1-NC and ESM1-mimic are subcutaneously injected into nude mice to generate tumors. ( B ) Statistics for tumor volume in nude mice. ( C ) Immunofluorescence double stain result plot.green:CA199, red:CD31; ( D ) Compared with the ESM1-NC group, CA199 and CD31 were significantly higher in the ESM1-mimic group. * p < 0.05, ** p < 0.01.

Article Snippet: CRC SW480 and SW620 cell lines and human normal colorectal cells (CCD-841CoN, IM-H477) were purchased from Nanjing KeyGen Biotech Co., Ltd. SW480 and SW620 cells were inoculated into L15 medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin and incubated in an incubator at 37°C with 5% CO 2 .

Techniques: Transfection, Injection, Immunofluorescence, Staining

Journal: Aging (Albany NY)

Article Title: ESM1 promotes angiogenesis in colorectal cancer by activating PI3K/Akt/mTOR pathway, thus accelerating tumor progression

doi: 10.18632/aging.204559

Figure Lengend Snippet: ESM1 shows an elevated expression in CRC cells and promotes the proliferation of CRC cells. ( A , B ) Relative protein expressions of p-PI3K, p-Akt, p-mTOR, MMP-2 and VEGF in ESM1-NC group, ESM1-mimic group, ESM1-NC + PI3K inhibitor group and ESM1-mimic + PI3K inhibitor group. ( C , D ) Expression of ESM1 in SW480 and SW620 cells. ( E ) Relative protein expression of ESM1 in CCD-841CoN, SW480 and SW620 cells. ** p < 0.01, ns: p > 0.05.

Article Snippet: CRC SW480 and SW620 cell lines and human normal colorectal cells (CCD-841CoN, IM-H477) were purchased from Nanjing KeyGen Biotech Co., Ltd. SW480 and SW620 cells were inoculated into L15 medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin and incubated in an incubator at 37°C with 5% CO 2 .

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: Alkannin-Induced Oxidative DNA Damage Synergizes With PARP Inhibition to Cause Cancer-Specific Cytotoxicity

doi: 10.3389/fphar.2020.610205

Figure Lengend Snippet: Sublethal doses of alkannin elevate ROS levels in colorectal cancer cells. (A) MTT assay. Cells were treated with 0.1, 0.2, 0.4, 0.8, 1.6, 2.4, 3.2, 4.8, 6.4, 9.6, 12.8, 19.2, 25.6, 38.4 or 51.2 μM alkannin for 24 h. The IC 50 values of alkannin against the SW480 and SW1116 colorectal cancer cell lines were calculated using the GraphPad Prism software. Data were shown as average ± SD from three independent experiments. (B) Colony formation assay. SW480 and SW1116 cancer cells were treated with alkannin at the indicated doses for 7 days and data from three independent experiments were presented as mean ± SD. (C) Representative images of DCFH-DA staining. SW480 cells were treated by alkannin at the indicated doses for 3 h (scale bars: 50 μm). (D) Measurement of ROS by flow cytometry. Treatment by 0.75 μM alkannin for 3 h induced a significant ROS increase in the SW480 cancer but not the NCM460 noncancerous cells. NAC suppressed the ROS increase in the cancer cells. (E) Quantification of flow cytometry measurements of ROS ( n = 3). n.s.: not significant, *: p < 0.05, ***: p < 0.001.

Article Snippet: 2 × 10 6 SW480 cells were inoculated subcutaneously into the flank of 8-week old male athymic BALB/c nude mice (Charles River, Boston, MA, United States), and the mice were randomly placed in control and treatment groups (5 animals per group).

Techniques: MTT Assay, Software, Colony Assay, Staining, Flow Cytometry

Journal: Frontiers in Pharmacology

Article Title: Alkannin-Induced Oxidative DNA Damage Synergizes With PARP Inhibition to Cause Cancer-Specific Cytotoxicity

doi: 10.3389/fphar.2020.610205

Figure Lengend Snippet: Sublethal alkannin-induced ROS elevation causes oxidative DNA damage. (A) Staining and measurement of cellular 8-oxoG. SW480 cells were treated by alkannin for 3 h and stained with Cy3-avidin (scale bars: 25 μm). Data from three independent experiments were presented as mean ± SD. (B) Alkaline comet assay. SW480 cells were treated by 0.75 μM alkannin for 3 h (scale bars: 25 μm). Tail moment was defined as percentage of tail DNA × tail length and was quantified using the TriTek CometScore software ( n = 3). (C) 53BP1 staining. SW480 cells were treated by 0.75 μM alkannin for the indicated times (scale bars: 10 μm). 53BP1 positive cells were quantified using the ImageJ software ( n = 3). n.s.: not significant, *: p < 0.05, **: p < 0.01, ***: p < 0.001.

Article Snippet: 2 × 10 6 SW480 cells were inoculated subcutaneously into the flank of 8-week old male athymic BALB/c nude mice (Charles River, Boston, MA, United States), and the mice were randomly placed in control and treatment groups (5 animals per group).

Techniques: Staining, Avidin-Biotin Assay, Alkaline Single Cell Gel Electrophoresis, Software

Journal: Frontiers in Pharmacology

Article Title: Alkannin-Induced Oxidative DNA Damage Synergizes With PARP Inhibition to Cause Cancer-Specific Cytotoxicity

doi: 10.3389/fphar.2020.610205

Figure Lengend Snippet: Sublethal alkannin sensitizes colorectal cancer cells to PARP-trapping. (A) SW480 cells readily formed RAD51 foci in response to 12 h of treatment by 5 μM SN-38, indicating that they were able to assemble recombination filaments normally (scale bars: 10 μm). (B) MTT assay. SW480 cells were treated with 0.125, 0.25, 0.5, 1.0, 2.0, 4.0, 8.0 or 16.0 μM olaparib alone or together with 0.75 μM alkannin with or without the OGG1 inhibitor O8 for 72 h. (C) Colony formation assay. SW480 and NCM460 cells were treated by the indicated drugs for 7 days (alkannin 0.75 μM, olaparib 10 μM, veliparib 10 μM, niraparib 5 μM, talazoparib 0.25 μM). Data from three independent experiments were presented as mean ± SD. (D) Determination of CI values. SW480 cells were treated by 1.5 or 0.75 μM alkannin combined with olaparib at the indicated concentrations (2, 4, 8, 16, 32, 64, 128 and 256 μM) for 72 h. (E,F) MTT assay. SW480 cells were treated with niraparib (E) or talazoparib (F) alone or together with 0.75 μM alkannin with or without the OGG1 inhibitor O8 for 72 h n.s.: not significant, ***: p < 0.001.

Article Snippet: 2 × 10 6 SW480 cells were inoculated subcutaneously into the flank of 8-week old male athymic BALB/c nude mice (Charles River, Boston, MA, United States), and the mice were randomly placed in control and treatment groups (5 animals per group).

Techniques: MTT Assay, Colony Assay

Journal: Frontiers in Pharmacology

Article Title: Alkannin-Induced Oxidative DNA Damage Synergizes With PARP Inhibition to Cause Cancer-Specific Cytotoxicity

doi: 10.3389/fphar.2020.610205

Figure Lengend Snippet: The combination of alkannin and olaparib induces intense replication stress and extensive DNA strand breaks. SW480 cells were treated by 0.75 μM alkannin, 10 μM olaparib or the combination of the two, with or without NAC, for 3 h. (A) Immunofluorescent staining of γH2AX (scale bars: 25 μm). Data from three independent experiments were presented as mean + SD. n.s.: not significant, ***: p < 0.001 vs vehicle control. (B) Alkaline comet assay and (C) 53BP1 staining ( n = 3). ***: p < 0.001, alkannin and olaparib combined vs alkannin or olaparib alone.

Article Snippet: 2 × 10 6 SW480 cells were inoculated subcutaneously into the flank of 8-week old male athymic BALB/c nude mice (Charles River, Boston, MA, United States), and the mice were randomly placed in control and treatment groups (5 animals per group).

Techniques: Staining, Control, Alkaline Single Cell Gel Electrophoresis

Journal: Frontiers in Pharmacology

Article Title: Alkannin-Induced Oxidative DNA Damage Synergizes With PARP Inhibition to Cause Cancer-Specific Cytotoxicity

doi: 10.3389/fphar.2020.610205

Figure Lengend Snippet: DDR activation leads to cell cycle arrest and apoptosis. (A) Western blot. SW480 cells were treated by 0.75 μM alkannin, 10 μM olaparib or the combination of the two, with or without NAC, for 3 h. (B) Flow cytometry analysis of cell cycle. SW480 cells were treated by 0.75 μM alkannin, 10 μM olaparib or the combination of the two for 48 h ( n = 3) (note: the subG 1 population was excluded from these measurements). (C) Measurement of mitochondrial membrane potential and (D) Western blot. SW480 cells were treated by the combination of 1.5 or 0.75 μM alkannin and 10 μM olaparib for 12 h. (E) Flow cytometry analysis of apoptosis. SW480 cells were treated by 0.75 μM alkannin, 10 μM olaparib or the combination of the two for 48 h ( n = 3). n.s.: not significant, *: p < 0.05, ***: p < 0.001.

Article Snippet: 2 × 10 6 SW480 cells were inoculated subcutaneously into the flank of 8-week old male athymic BALB/c nude mice (Charles River, Boston, MA, United States), and the mice were randomly placed in control and treatment groups (5 animals per group).

Techniques: Activation Assay, Western Blot, Flow Cytometry, Membrane