Journal: Cancer research

Article Title: Inhibition of Cholinergic Signaling Causes Apoptosis in Human Bronchioalveolar Carcinoma

doi: 10.1158/0008-5472.CAN-12-3190

Figure Lengend Snippet: Human BACs and normal HPAEpiCs express cholinergic proteins. A, ELISA assays show that human BAC cell lines and HPAEpiCs express multiple nAChR subunits. The assay was completed in duplicate, and the whole experiment was carried out 2 independent times. Results indicated by a different letter are significantly different (P < 0.05). B, HPAEpiCs and human BACs express AChE, ChAT, CHT1, and VAChT. H520 human SCC-L cells were used as the positive control for the experiments outlined in A and B. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as the loading control, and the results were quantitated by densitometric analysis. The experiment was repeated twice, and the representative data are shown. C and D, immunohistochemistry of the human BAC microarray showed that BAC tumors (isolated from patients) express ChAT and VAChT in the cytoplasm, adjacent to the hematoxylin-counterstained dark nuclei in the tissue samples. A tumor microarray containing 81 cores of human BACs was used for these experiments and 4 panels of representative photos are shown. Scale bar, 20 μm.

Article Snippet: Immunohistochemical staining of VAChT, ChAT in human BAC tissue microarray and normal lung TMA Human BAC tissue microarray (TMA) slides (Abnova) were deparaffinized and rehydrated as described previously ( 19 , 20 ).

Techniques: Enzyme-linked Immunosorbent Assay, Positive Control, Control, Immunohistochemistry, Microarray, Isolation

Journal: Frontiers in Oncology

Article Title: PGC1β Regulates Breast Tumor Growth and Metastasis by SREBP1-Mediated HKDC1 Expression

doi: 10.3389/fonc.2019.00290

Figure Lengend Snippet: HKDC1 expression involves with tumor growth and metastasis in vivo . (A,B) Tissue Microarray slides (#CHTN BrCaProg1) were obtained from CHTN (Cooperative Human Tissue Network), and the immunohistochemistry was performed using a primary rabbit antibody for either PGC1β or HKDC1 and a second anti-rabbit- FITC. Sixty cells from normal breast tissue (normal), primary tumor (tumor), and metastatic (metastasis) tissues were quantitated by Image J. (A) Protein quantitation, n = 3. *, P < 0.05, vs. Normal group; ¶, P < 0.05 vs. Tumor group. (B) Representative pictures for (A) . (C–H) The nude mice were injected with treated MCF7 cells through the tail vein for in vivo xenograft tumor development study, and the treated mice were sacrificed for further analysis. (C) The tumor tissues from the lung were isolated for mRNA analysis by qPCR, n = 4. *, P < 0.05, vs. CTL group. (D) Superoxide anion release from tumor tissues, n = 5, *, P < 0.05, vs. CTL group; #, P < 0.05, vs. shPGC1β group. (E–G) Mice were killed upon 20% weight loss, and the lungs were harvested for terminal analysis. The metastatic tumor nodules from the lungs were counted, and then the formalin-fixed, paraffin-embedded tumor tissue of the lung was sectioned to 4 mm thickness, and the histopathological analyses were performed with H&E staining. Images were taken using a Carl Zeiss MIRAX MIDI slide scanner, and the lung tumor spots were analyzed using a 3DHISTECH Pannoramic Viewer. (E) Tumor colony formation in lung, n = 9. *, P < 0.05, vs. CTL group; ¶, P < 0.05, vs. ↑PGC1β group; #, P < 0.05, vs. shPGC1β group. (F) Quantitated lung tumor spots, n = 5. *, P < 0.05, vs. CTL group; ¶, P < 0.05, vs. ↑PGC1β group; #, P < 0.05, vs. shPGC1β group. (G) Representative picture by H&E staining. (H) Kaplan-Meier analysis comparing survival of mice between each treatment group, P -value represents log-rank Mantel-Cox test result, n = 9. Results are expressed as mean ± SEM.

Article Snippet: CHTN BrCaProg1 Microarray slides were obtained from CHTN, including normal, tumor, and metastatic breast cancer tissues.

Techniques: Expressing, In Vivo, Microarray, Immunohistochemistry, Protein Quantitation, Injection, Isolation, Formalin-fixed Paraffin-Embedded, Staining