Journal: Scientific Reports

Article Title: PIK75 effectively reverses PI3K‒AKT activation caused by palbociclib resistance and synergistically inhibits the progression of esophageal squamous cell carcinoma

doi: 10.1038/s41598-025-26858-5

Figure Lengend Snippet: Palbociclib unexpectedly activates the PI3K-AKT pathway which is associated with the resistance to CDK inhibitor. ( A ) Top 20 KEGG-enriched pathways of differential genes down-regulated by addition of palbociclib in KYSE30 cell. ( B ) GO analysis of differential genes down-regulated by addition of palbociclib in KYSE30 cells. ( C ) RT-qPCR analysis of mRNA levels of CCNE1, CDC25A and CDC6 in negative control and ESCC cells after addition of 20 µM palbociclib. ( D ) Western blot analysis and statistical plots of CCNE1, CDC25A, CDC6 and GAPDH in negative control and ESCC cells after addition of 20 µM palbociclib. ( E ) Top 15 KEGG-enriched pathways of differential genes up-regulated by addition of palbociclib in KYSE30 cell. ( F ) Western blot analysis of PIK3CA, p-AKT, AKT and GAPDH in negative control and ESCC cells after addition of 20 µM palbociclib. The Data shown represent the mean ± SD.

Article Snippet: Western blotting was performed with overnight incubation with primary antibodies against GAPDH (CST, USA), p-AKT (CST, USA), AKT (CST, USA), CDC25A (Proteintech), CCNE1 (Proteintech), CDC6 (Proteintech), and PIK3CA (Proteintech).

Techniques: Quantitative RT-PCR, Negative Control, Western Blot

Journal: Scientific Reports

Article Title: PIK75 effectively reverses PI3K‒AKT activation caused by palbociclib resistance and synergistically inhibits the progression of esophageal squamous cell carcinoma

doi: 10.1038/s41598-025-26858-5

Figure Lengend Snippet: PIK75 inhibits the activation of the PI3K-AKT pathway and promotes cell apoptosis in ESCC cells. ( A ) Representative images of colony growth in different ESCC cells with the addition of PIK75, n = 3. ( B ) Cell proliferation assay after addition of PIK75 to in different ESCC cell lines, n = 3 ( C ) Representative images for the detection of apoptosis by flow technology, n = 3. ( D ) Top 15 KEGG-enriched pathways of differential genes by addition of PIK75 in KYSE30 cell. ( E ) GO analysis of differential genes by addition of PIK75 in KYSE30 cell. ( F ) RT-qPCR analysis of mRNA levels of CCNE1, CDC25A and CDC6 in negative control and ESCC cells after addition of 5 µM PIK75. (G-H) Western blot analysis of PIK3CA, AKT, p-AKT, CCNE1, CDC25A, CDC6 and GAPDH in negative control and ESCC cells after addition of 5 µM PIK75. The Data shown represent the mean ± SD.

Article Snippet: Western blotting was performed with overnight incubation with primary antibodies against GAPDH (CST, USA), p-AKT (CST, USA), AKT (CST, USA), CDC25A (Proteintech), CCNE1 (Proteintech), CDC6 (Proteintech), and PIK3CA (Proteintech).

Techniques: Activation Assay, Proliferation Assay, Quantitative RT-PCR, Negative Control, Western Blot

Journal: Scientific Reports

Article Title: PIK75 effectively reverses PI3K‒AKT activation caused by palbociclib resistance and synergistically inhibits the progression of esophageal squamous cell carcinoma

doi: 10.1038/s41598-025-26858-5

Figure Lengend Snippet: Phosphorylation of AKT is responsible for resistance to CDK inhibitor. ( A ) Vene plots of the three gene sets for genes up-regulated with palbociclib, genes down-regulated with PIK75, and genes down-regulated with the combination of drugs in KYSE30 cell. ( B ) Top 15 KEGG-enriched pathways of intersecting differential genes screened by Vene plots in KYSE30 cell. ( C ) RT-qPCR analysis of mRNA levels of CCNE1, CDC25A and CDC6 in negative control and ESCC cells after addition of PIK75 or/and Palbociclib. ( D ) Western blot analysis of PIK3CA, AKT, p-AKT, CCNE1, CDC25A, CDC6 and GAPDH in negative control and ESCC cells after addition of palbociclib or/and PIK75. # p < 0.05, ## p < 0.01, ### p < 0.001 vs. NC group. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Combine group. The Data shown represent the mean ± SD.

Article Snippet: Western blotting was performed with overnight incubation with primary antibodies against GAPDH (CST, USA), p-AKT (CST, USA), AKT (CST, USA), CDC25A (Proteintech), CCNE1 (Proteintech), CDC6 (Proteintech), and PIK3CA (Proteintech).

Techniques: Phospho-proteomics, Quantitative RT-PCR, Negative Control, Western Blot

Journal: Scientific Reports

Article Title: PIK75 effectively reverses PI3K‒AKT activation caused by palbociclib resistance and synergistically inhibits the progression of esophageal squamous cell carcinoma

doi: 10.1038/s41598-025-26858-5

Figure Lengend Snippet: PIK75 synergizes with palbociclib to inhibit ESCC tumor progression. ( A ) Representative images of the tumor, n = 7. ( B ) Tumor volume growth curves for different groups, n = 7. ( C ) Tumor weights of different groups, n = 7. ( D ) Percentage change in body weight of mice during tumor growth, n = 7. ( E ) Representative images of H&E staining, KI67-positive cell staining, and Tunel staining of paraffin sections from different groups of mouse tumors, n = 3. ( F ) Statistical chart of staining of Ki-67 positive cells, n = 3. ( G ) Staining statistics of TUNEL-positive cells, n = 3. ( H - I ) Western blot analysis of PIK3CA, AKT, p-AKT, CCNE1, CDC25A, CDC6 and GAPDH in tumor tissues of different groups of mice, n = 4–7. ( J ) RT-qPCR analysis of mRNA levels of CCNE1, CDC25A and CDC6 in tumor tissues of different groups of mice, n = 4–7. # p < 0.05, ## p < 0.01, ### p < 0.001 vs. NC group mice. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Combine group mice. The Data shown represent the mean ± SD.

Article Snippet: Western blotting was performed with overnight incubation with primary antibodies against GAPDH (CST, USA), p-AKT (CST, USA), AKT (CST, USA), CDC25A (Proteintech), CCNE1 (Proteintech), CDC6 (Proteintech), and PIK3CA (Proteintech).

Techniques: Staining, TUNEL Assay, Western Blot, Quantitative RT-PCR