Journal: International Journal of Oncology

Article Title: miR-150 inhibits proliferation and tumorigenicity via retarding G1/S phase transition in nasopharyngeal carcinoma

doi: 10.3892/ijo.2017.3909

Figure Lengend Snippet: The primers used in the reactions for real-time RT-PCR.

Article Snippet: Small interfering RNA (siRNA) for CCND1, CCND2, CDK2 and CCNE2 knockdown were obtained from Santa Cruz (Dallas, TX, USA).

Techniques: Sequencing

Journal: International Journal of Oncology

Article Title: miR-150 inhibits proliferation and tumorigenicity via retarding G1/S phase transition in nasopharyngeal carcinoma

doi: 10.3892/ijo.2017.3909

Figure Lengend Snippet: The primers used in the reactions for clone PCR.

Article Snippet: Small interfering RNA (siRNA) for CCND1, CCND2, CDK2 and CCNE2 knockdown were obtained from Santa Cruz (Dallas, TX, USA).

Techniques: Sequencing

Journal: International Journal of Oncology

Article Title: miR-150 inhibits proliferation and tumorigenicity via retarding G1/S phase transition in nasopharyngeal carcinoma

doi: 10.3892/ijo.2017.3909

Figure Lengend Snippet: miR-150 inhibits the cell cycle of nasopharyngeal carcinoma cells. (A) Flow cytometric analysis of the indicated nasopharyngeal carcinoma cells. Each bar represents the mean values ± SD of three independent experiments. * P<0.05. (B) Real-time PCR analysis of CCND1, CCND2, CKD2 and CCNE2 expression in the indicated cells. Transcript levels were normalized by GAPDH expression. Each bar represents the mean values ± SD of three independent experiments. * P<0.05. (C) Western blot analysis of CCND1, CCND2, CDK2, CCNE2 and p-Rb expression in the indicated cells. (D) Real-time PCR analysis of CDK4, CDK6, CCND3 and CCNE2 mRNA expression in overexpressed miR-150 and anti-miR-150 NPC cells, and their respective control vectors. U6 was used as the loading control. Each bar represents the mean values ± SD of three independent experiments. * P<0.05. (E and F) Relative Rb activity and E2F reporter activity in the indicated cells. Each bar represents the mean values ± SD of three independent experiments. * P<0.05.

Article Snippet: Small interfering RNA (siRNA) for CCND1, CCND2, CDK2 and CCNE2 knockdown were obtained from Santa Cruz (Dallas, TX, USA).

Techniques: Real-time Polymerase Chain Reaction, Expressing, Western Blot, Control, Activity Assay

Journal: International Journal of Oncology

Article Title: miR-150 inhibits proliferation and tumorigenicity via retarding G1/S phase transition in nasopharyngeal carcinoma

doi: 10.3892/ijo.2017.3909

Figure Lengend Snippet: miR-150 inhibits the cell cycle progression by directly targeting CCND1, CCND2, CDK2 and CCNE2. (A) MiRNP IP (RIP) assay showing the association between miR-150 and CCND1, CCND2, CDK2 and CCNE2 transcripts in CNE-2 and HONE1 cells. Pulldown of IgG antibody served as the negative control. (B) RIP analysis show that CDK4, CDK6, CCND3 and CCNE1 were not affected by miR-150. RIP analysis, as assessed by immunoprecipitation of Ago2 in the indicated cells. IgG immunoprecipitation was used as a negative control. * P<0.05. (C) Predicted miR-150 targeting sequence in 3′-UTRs of CCND1, CCND2, CDK2 and CCNE2. (D) Luciferase assay of cells transfected with pmirGLO-3′-UTR reporter of CCND1, CCND2, CDK2 and CCNE2 in miR-150 overexpressing and silencing in CNE-2 and HONE1 cells, respectively. (E) Individual silencing of CCND1, CCND2, CDK2 and CCNE2 increased the percentage of NPC cells in G 0 phase, while the percentage of cells in S phase was decreased in miR-150-silencing cells.

Article Snippet: Small interfering RNA (siRNA) for CCND1, CCND2, CDK2 and CCNE2 knockdown were obtained from Santa Cruz (Dallas, TX, USA).

Techniques: Negative Control, Immunoprecipitation, Sequencing, Luciferase, Transfection

Journal: International Journal of Oncology

Article Title: miR-150 inhibits proliferation and tumorigenicity via retarding G1/S phase transition in nasopharyngeal carcinoma

doi: 10.3892/ijo.2017.3909

Figure Lengend Snippet: CCND1, CCND2, CDK2 and CCNE2 mediates the miR-150-downregulation-induced NPC cell growth. (A) Individual silencing of CCND1, CCND2, CDK2 and CCNE2 rescued the proliferation rate enhanced by anti-miR-150. (B) Individual silencing of CCND1, CCND2, CDK2 and CCNE2 rescued the colony-formation ability enhanced by anti-miR-150. (C) Individual silencing of CCND1, CCND2, CDK2 and CCNE2 rescued the anchorage-independent growth ability enhanced by anti-miR-150.

Article Snippet: Small interfering RNA (siRNA) for CCND1, CCND2, CDK2 and CCNE2 knockdown were obtained from Santa Cruz (Dallas, TX, USA).

Techniques:

Journal: International Journal of Oncology

Article Title: miR-150 inhibits proliferation and tumorigenicity via retarding G1/S phase transition in nasopharyngeal carcinoma

doi: 10.3892/ijo.2017.3909

Figure Lengend Snippet: Clinical relevance of miR-150 with CCND1, CCND2, CDK2 and CCNE2 in nasopharyngeal carcinoma tissues. (A) miR-150 expression in eight freshly-frozen human nasopharyngeal carcinoma tissues. Transcript levels were normalized by U6 expression. One tissue sample was randomly selected and numbered as T1, other 7 tissue samples were numbered in order as T2, T3, T4, T5, T6, T7 and T8. The expression levels of miR-150 in each NPC tissue were normalized by the corresponding miR-150 expression in T1 NPC tissue. The relative expression levels of miR-150 after normalization were used to perform the correlation analysis among miR-150 and CCND1, CCND2, CKD2 and CCNE2 expression. (B) Analysis of CCND1, CCND2, CDK2 and CCNE2 expression in eight freshly-frozen human nasopharyngeal carcinoma tissues. Transcript levels were normalized by GAPDH expression. One tissue sample was randomly selected and numbered as T1, other 7 tissue samples were numbered in order as T2, T3, T4, T5, T6, T7 and T8. The expression levels of CCND1, CCND2, CKD2 and CCNE2 in each NPC tissue were normalized by the corresponding CCND1, CCND2, CKD2 and CCNE2 expression levels in T1 NPC tissue. The relative expression levels of CCND1, CCND2, CKD2 and CCNE2 after normalization were used to perform the correlation analysis among miR-150 and CCND1, CCND2, CKD2 and CCNE2 expression. (C) Correlation between miR-150 levels and CCND1, CCND2, CDK2 and CCNE2 expression in nasopharyngeal carcinoma tissues.

Article Snippet: Small interfering RNA (siRNA) for CCND1, CCND2, CDK2 and CCNE2 knockdown were obtained from Santa Cruz (Dallas, TX, USA).

Techniques: Expressing