Fig. 7.

TERRA is overexpressed in human tumor tissues. (A) Northern blot analysis of TERRA RNA isolated from normal ovarian tissue, advanced primary ovarian cancer and metastatic ovarian cancer tissue. Numbers at the bottom show the average value of TERRA signals relative to 18S RNA signals in tumor versus normal tissues from two independent northern blots. Asterisks indicate that the sample is not included in the analysis because of the degradation of the 18S signal. (B) Quantification of average TERRA levels from many northern blot analyses using RNA isolated from tissues derived from two normal ovaries (540071 and 709152), eight primary and nine metastatic ovarian cancers, a representative of which is shown in A. Bar graph represents TERRA signal intensity relative to the 18S signal, and average relative intensity for two normal ovary tissues was set at 100. (C) qRT-PCR analysis of Ki-67 expression and TERRA levels in the indicated ovarian cancers and normal ovary tissue (709152). TERRA levels were assayed using primers specific for TERRA RNA transcribed from subtelomeres of human chromosome 2q, 10q, 13q, as indicated. ΔΔCT methods relative to normal ovary and Gapdh were used to calculate relative RT-PCR between normal and tumor samples. Bar graph represents the average value from three independent PCR reactions (means ± s.d.). (D) The same as in C, except using primers specific for TERRA RNA transcribed from subtelomeres of human chromosome XqYq, 15q, 16p, as indicated. (E) TERRA expression in dissected primary human tumor tissue from carcinoma of the stomach, lung and colon (normal and tumor) analyzed by northern blotting. 18S RNA expression is shown as quantification control. Equal intensity of 18S* signals indicate that RNA from each sample were subject to similar levels of degradation during the preparation process. The same northern blot was stripped, and reprobed with ³²P-labeled (TTAGGG)₄ probe for anti-sense TERRA (right panel). Numbers on the left show the position of markers (in kb). Numbers at the bottom show the value of TERRA signals relative to the 18S RNA signals in tumor versus normal tissues. (F) qRT-PCR analysis of Ki-67 expression (left panel) and TERRA levels (right panel) in the tumor and patient matched control tissues from stomach. TERRA levels were assayed using primers specific for TERRA RNA transcribed from subtelomeres of human chromosome 2q, 17q, 10q, 13q, 15q, 16q and XqYq, as indicated. Bar graph represents the average value from at least three independent PCR reactions (means ± s.d.). (G) The same as in F, except in the tumor and patient matched control tissues from lung. (H) The same as in F, except in the tumor and patient matched control tissues from the colon.