Journal: Cancer cell international

Article Title: Interplay between lncRNA RP11-367G18.1 variant 2 and YY1 plays a vital role in hypoxia-mediated gene expression and tumorigenesis.

doi: 10.1186/s12935-023-03067-6

Figure Lengend Snippet: Fig. 2 RP11-367G18.1 variant 2 interacts with YY1 to activate H4K16Ac. (A) RNA pull-down assay and Coomassie blue staining revealed 11 RP11-367G18.1 variant 2-specific bands (left). Protein identity of the 11 bands was analyzed via LC-MS/MS and was shown in a table (right). (B) Knockdown of YY1 sup pressed H4K16Ac levels in H1299 cells. (C) Overexpression of YY1 increased H4K16Ac levels in H1299 cells. (D) YY1 was pulled down by biotinylated sense RP11-367G18.1 variant 2. Beads or biotinylated anti-sense RP11-367G18.1 variant 2 were used as the negative control. (E) YY1-induced H4K16Ac activation was suppressed following RP11-367G18.1 variant 2 knockdown. (F) mRNA levels of YY1 were upregulated under hypoxia in FADU and MCF7 cells. (G) Protein levels of YY1 were upregulated under hypoxia in FADU and MCF7 cells. (H) Reporter constructs containing wild-type and mutant HREs in YY1 promoter were shown (left). Reporter assay revealed that HRE (-493/-489) was responsive to hypoxia in YY1 promoter (right). (I) ChIP assay revealed that HIF-1α bound to the YY1 proximal promoters containing HRE under hypoxia. For hypoxic conditions, cells were cultured in 1% O2, 5% CO2, and 94% N2 for 18 h. Scr, Scrambled; Cont, control; V2, variant 2. Data are represented as the mean ± SD. *P < 0.05

Article Snippet: Human cell lines with low (MCF7 breast cancer and FADU HNSC cell lines) and high (H1299 non-small lung cancer and MDA-MB-231 breast cancer cell lines; Figure S1A) HIF-1α levels were purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Variant Assay, Pull Down Assay, Staining, Liquid Chromatography with Mass Spectroscopy, Knockdown, Over Expression, Negative Control, Activation Assay, Construct, Mutagenesis, Reporter Assay, Cell Culture, Control

Journal: Cancer cell international

Article Title: Interplay between lncRNA RP11-367G18.1 variant 2 and YY1 plays a vital role in hypoxia-mediated gene expression and tumorigenesis.

doi: 10.1186/s12935-023-03067-6

Figure Lengend Snippet: Fig. 3 RP11-367G18.1 variant 2-YY1 complex activates H4K16Ac. (A) Hypoxia enhanced H4K16Ac activation. (B) Immunofluorescence staining revealed that RP11-367G18.1 variant 2 was colocalized with YY1 and H4K16Ac in FADU cells under hypoxia. (C) Overexpression of RP11-367G18.1 variant 2 and YY1 enhanced H4K16Ac activation. For hypoxic conditions, cells were cultured in 1% O2, 5% CO2, and 94% N2 for 18 h

Article Snippet: Human cell lines with low (MCF7 breast cancer and FADU HNSC cell lines) and high (H1299 non-small lung cancer and MDA-MB-231 breast cancer cell lines; Figure S1A) HIF-1α levels were purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Variant Assay, Activation Assay, Immunofluorescence, Staining, Over Expression, Cell Culture

Journal: Cancer cell international

Article Title: Interplay between lncRNA RP11-367G18.1 variant 2 and YY1 plays a vital role in hypoxia-mediated gene expression and tumorigenesis.

doi: 10.1186/s12935-023-03067-6

Figure Lengend Snippet: Fig. 4 RP11-367G18.1 and YY1 are associated with worse outcomes in patients with HNSC. (A and B) Expression levels of RP11-367G18.1 and YY1 were higher in tumor tissues than those in normal tissues in patients with HNSC from TCGA dataset. (C) Positive correlation between RP11-367G18.1 and YY1 in HNSC tissues was shown. (D) Patients with HNSC with high expression levels of RP11-367G18.1 and YY1 showed the worse overall survival. (E) Patients with HNSC with high expression levels of RP11-367G18.1, YY1, and HIF-1α showed the worse overall survival

Article Snippet: Human cell lines with low (MCF7 breast cancer and FADU HNSC cell lines) and high (H1299 non-small lung cancer and MDA-MB-231 breast cancer cell lines; Figure S1A) HIF-1α levels were purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Expressing

Journal: Cancer cell international

Article Title: Interplay between lncRNA RP11-367G18.1 variant 2 and YY1 plays a vital role in hypoxia-mediated gene expression and tumorigenesis.

doi: 10.1186/s12935-023-03067-6

Figure Lengend Snippet: Fig. 5 RP11-367G18.1 variant 2–YY1 complex mediates hypoxia-induced EMT. (A and B) Knockdown of RP11-367G18.1 variant 2 or YY1 suppressed EMT, cell migration, and invasion of FADU and MCF7 cells overexpressing HIF-1α (ΔODD). (C and D) Knockdown of RP11-367G18.1 variant 2 or YY1 suppressed EMT, cell migration, and invasion of FADU and MCF7 cells under hypoxia. Glut1 and LDHA, the hypoxia-inducible genes, served as positive control. For hypoxic conditions, cells were cultured in 1% O2, 5% CO2, and 94% N2 for 18 h. Scr, Scrambled; Cont, control; V2, variant 2; N, normoxia; H, hypoxia. Data are represented as the mean ± SD. *P < 0.05

Article Snippet: Human cell lines with low (MCF7 breast cancer and FADU HNSC cell lines) and high (H1299 non-small lung cancer and MDA-MB-231 breast cancer cell lines; Figure S1A) HIF-1α levels were purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Variant Assay, Knockdown, Migration, Positive Control, Cell Culture, Control

Journal: Cancer cell international

Article Title: Interplay between lncRNA RP11-367G18.1 variant 2 and YY1 plays a vital role in hypoxia-mediated gene expression and tumorigenesis.

doi: 10.1186/s12935-023-03067-6

Figure Lengend Snippet: Fig. 6 RP11-367G18.1 variant 2–YY1 complex is essential for hypoxia-induced tumorigenicity. (A) Knockdown of YY1 decreased the colony formation of FADU cells overexpressing RP11-367G18.1 variant 2. (B) Knockdown of RP11-367G18.1 variant 2 suppressed the colony formation of FADU cells over expressing YY1. (C) Knockdown of RP11-367G18.1 variant 2 or YY1 suppressed the colony formation of H1299 cells overexpressing HIF-1α (ΔODD). (D) Knockdown of RP11-367G18.1 variant 2 or YY1 suppressed HIF-1α (ΔODD)-enhanced tumor volume of FADU cell-derived xenografts. Scr, Scrambled; Cont, control; V2, variant 2. Data are represented as the mean ± SD. *P < 0.05

Article Snippet: Human cell lines with low (MCF7 breast cancer and FADU HNSC cell lines) and high (H1299 non-small lung cancer and MDA-MB-231 breast cancer cell lines; Figure S1A) HIF-1α levels were purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Variant Assay, Knockdown, Expressing, Derivative Assay, Control

Journal: Cancer cell international

Article Title: Interplay between lncRNA RP11-367G18.1 variant 2 and YY1 plays a vital role in hypoxia-mediated gene expression and tumorigenesis.

doi: 10.1186/s12935-023-03067-6

Figure Lengend Snippet: Fig. 7 RP11-367G18.1 variant 2–YY1 complex regulates gene expression. (A) The transcript expression levels of RP11-367G18.1 variant 2, YY1, HK2, TGFBI, VEGFC, and LIF were measured at the indicated time points (in hours) under hypoxic conditions in FADU cells. (B) The proteins expressions levels of HIF- 1α, YY1 and H4K16Ac were measured at the indicated time points (in hours) under hypoxic conditions in FADU cells. (C) Knockdown of RP11-367G18.1 variant 2 or YY1 inhibited the expression of HK2, TGFBI. VEGFC, and LIF under hypoxia in FADU cells. (D) Knockdown of RP11-367G18.1 variant 2 suppressed the level of H4K16Ac and the binding of YY1 to the proximal promoters of HK2, TGFBI, VEGFC, and LIF under hypoxic conditions. Glut1, a hypoxia-inducible gene, served as positive control. For hypoxic conditions, cells were cultured in 1% O2, 5% CO2, and 94% N2 for 18 h. Cont, control; V2, variant 2. Data are represented as the mean ± SD. Student’s t-test, *P < 0.05

Article Snippet: Human cell lines with low (MCF7 breast cancer and FADU HNSC cell lines) and high (H1299 non-small lung cancer and MDA-MB-231 breast cancer cell lines; Figure S1A) HIF-1α levels were purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Variant Assay, Gene Expression, Expressing, Knockdown, Binding Assay, Positive Control, Cell Culture, Control