Journal: Cancer Medicine

Article Title: Comprehensive analysis indicated that NDE1 is a potential biomarker for pan‐cancer and promotes bladder cancer progression

doi: 10.1002/cam4.6931

Figure Lengend Snippet: Biological functions of NDE1 in BLCA. (A, C) The knockdown efficiency of NDE1 in T24 cell line was verified by western blot; (B) and (D) NDE1 knockdown efficiency in 5637 cell line was detected by western blot; (E, F) The effect of NDE1 on the migration and invasion of T24 cells was investigated by scratch assay; (G, H) The effect of NDE1 on the migration and invasion of 5637 cells was investigated by scratch assay. (I) To investigate the effect of NDE1 on the proliferation of T24 cells by CCK‐8 assay; (J) CCK‐8 assay was used to investigate the effect of NDE1 on the proliferation of 5637 cells; (K, L) transwell assay was used to investigate the effect of NDE1 on the migration and invasion of T24 and 5637 cells. (M, N) The effect of NDE1 on the proliferation ability of T24 and 5637 cells was explored by colony formation assay. * p < 0.05; ** p < 0.01; *** p < 0.001 and **** p < 0.0001.

Article Snippet: Procell Life Science and Technology sold the T24 and 5637 cell lines for human bladder urothelial cancer.

Techniques: Knockdown, Western Blot, Migration, Wound Healing Assay, CCK-8 Assay, Transwell Assay, Colony Assay

Journal: Cancer Medicine

Article Title: Comprehensive analysis indicated that NDE1 is a potential biomarker for pan‐cancer and promotes bladder cancer progression

doi: 10.1002/cam4.6931

Figure Lengend Snippet: Flow cytometry was used to detect the early apoptosis and late apoptosis cell population in T24 and 5637.

Article Snippet: Procell Life Science and Technology sold the T24 and 5637 cell lines for human bladder urothelial cancer.

Techniques: Flow Cytometry

Journal: Journal of Oncology

Article Title: Loss of Lactate/Proton Monocarboxylate Transporter 4 Induces Ferroptosis via the AMPK/ACC Pathway and Inhibition of Autophagy on Human Bladder Cancer 5637 Cell Line

doi: 10.1155/2023/2830306

Figure Lengend Snippet: MCT4 was positively associated with poor prognosis in bladder cancer patients. Among 402 bladder cancer patients, the survival rate of 201 patients with high expression of MCT4 was significantly lower than that of 201 patients with low expression of MCT4. p (HR) was 0.023, and HR was 1.4. (b) The knockdown efficiency of MCT4 mRNA was detected by qRT-PCR. GAPDH was used as an internal reference. (c) Western Blot assay was used to detect the knockdown efficiency of MCT4. Tubulin was used as an internal control. (d) Clone formation assay was performed to detect the clone formation ability of bladder cancer cells after knockdown of MCT4. (e) Subcutaneous tumorigenicity of 5637 cells in nude mice after knockdown of MCT4.

Article Snippet: Transfected 5637 cells (600 × 10 4 cells/100 μ l) were then injected into the right axilla of 6-week-old male nude mice ( n = 12; Charles River, Beijing, China).

Techniques: Expressing, Knockdown, Quantitative RT-PCR, Western Blot, Control, Tube Formation Assay

Journal: Journal of Oncology

Article Title: Loss of Lactate/Proton Monocarboxylate Transporter 4 Induces Ferroptosis via the AMPK/ACC Pathway and Inhibition of Autophagy on Human Bladder Cancer 5637 Cell Line

doi: 10.1155/2023/2830306

Figure Lengend Snippet: Bladder cancer cells underwent ferroptosis after the knockdown of MCT4. (a) Pathway analysis showed that ferroptosis and autophagy-related pathways were activated after the knockdown of MCT4. (b) Western Blot assay detected the expression levels of FSP1 and SLC7A11 proteins. (c) The intracellular morphology was evaluated by TEM. (d) DCFH-DA probe detected ROS in 5637 cells in knockdown of the MCT4 group and control group after induction with 2.5 μ M RSL3 (APExBIO) for 24 h. Scale bar = 200 μ m. (e) MDA assay detected the changes of lipid metabolism disorder in knockdown of the MCT4 group and control group after 24 h induction with 2.5 μ M RSL3 (APExBIO). (f) LPO assay detected the level of lipid peroxidation in 5637 cells after knockdown of MCT4, and fer-1 (APExBIO) was a specific inhibitor of ferroptosis. Scale bar = 50 μ m. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗∗ p < 0.001.

Article Snippet: Transfected 5637 cells (600 × 10 4 cells/100 μ l) were then injected into the right axilla of 6-week-old male nude mice ( n = 12; Charles River, Beijing, China).

Techniques: Knockdown, Western Blot, Expressing, Control, Multiple Displacement Amplification

Journal: Journal of Oncology

Article Title: Loss of Lactate/Proton Monocarboxylate Transporter 4 Induces Ferroptosis via the AMPK/ACC Pathway and Inhibition of Autophagy on Human Bladder Cancer 5637 Cell Line

doi: 10.1155/2023/2830306

Figure Lengend Snippet: Autophagy in bladder cancer cells was inhibited after knockdown of MCT4. (a) Western Blot assay was used to detect the changes in the protein expression levels of autophagy-related proteins, including LC3A/B I and LC3A/B II in 5637 cells with MCT4 knockdown and the control group. Vinculin was used as an internal control. (b) Fluorescence microscopy (OLYMBUS IX83, Japan) detected that autophagy was inhibited in 5637 cells after transient transfection of siMCT4 for 48 h by AdPlus-mCherry-GFP-LC3B. Scale bar = 20 μ m. (c) Flow cytometry detected the changes of apoptosis in the knockdown group and the control group after treatment of 1 μ M CQ (Selleck) for 24 h. (d) Changes of caspase-Glo3/7 in the knockdown of the MCT4 group and the control group after treatment of 1 μ M CQ for 24 h were detected by a fluorescence microplate reader. ∗∗ p = 0.004.

Article Snippet: Transfected 5637 cells (600 × 10 4 cells/100 μ l) were then injected into the right axilla of 6-week-old male nude mice ( n = 12; Charles River, Beijing, China).

Techniques: Knockdown, Western Blot, Expressing, Control, Fluorescence, Microscopy, Transfection, Flow Cytometry