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. 2019 Dec 18;12(1):10. doi: 10.3390/cancers12010010

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© 2019 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Modulation effect of PTEN on maspin expression in bladder carcinoma cells. (A) The mock-knockdown RT4 (RT4_shCOL), PTEN-knockdown RT4 (RT4_shPTEN), mock-transfected T24 (T24-DNA), and PTEN-overexpressed T24 (T24-PTEN) cells were lysed, and the protein levels of maspin and PTEN were determined through immunoblot assays. (B) The cell proliferation of RT4_shCOL and RT4_shPTEN was determined through the ³H-thymidine incorporation assays (±SE, n = 4). (C) Four-week-old male athymic nude mice were divided randomly into two groups. Cells (5 × 10⁶) were injected subcutaneously in the dorsal area of the mice (n = 6), and tumors derived from RT4_shCOL and RT4_shMapsin cells were measured after sacrifice. (D) The tumor volumes (±SE, n = 6) were measured every week during the indicated period. The expressions of PTEN, Akt, pAkt^S473, pAkt^T308, and maspin in RT4, RT4-shCOL, and RT4-shPTEN cells (E) as well as in VO-OHpic-treated RT4 cells (F) were determined through immunoblot assays. The mRNA ratios of PTEN and maspin between (G) RT4_shCOL and RT4_shPTEN cells and between (H) T24-DNA and T24-Maspin cells (±SE, n = 4) were determined by RT-qPCR assays. (I) The expressions of Akt, pAkt^S473, pAkt^T308, maspin, and β-Actin in MK2206-treated T24 cells were determined through immunoblot assays. (J) The reporter activity of maspin reporter vector cotransfected with various dosages of PTEN, as indicated in the HT1376 cells. Data are expressed as the mean percentage ± SE of luciferase activity relative to the mock-transfected group (n = 6). * p < 0.05, ** p < 0.01. The numbers indicate the ratio of the target gene/β-Actin, pAkt^S473/Akt, or pAkt^T308/Akt in relation to RT-4, or T24 cells.