Abdulkareem Imran Haruna, Blair Maria
Department of Trauma and Orthopaedics Surgery, Leeds University Teaching Hospitals, Leeds, West Yorkshire, United Kingdom.
Niger Med J. 2013 Mar;54(2):100-6. doi: 10.4103/0300-1652.110041.
Previous studies reported that Non-steroidal Anti-inflammatory Drugs (NSAIDs), chemicals, and food supplements can be used to up-regulate the PTEN mRNA and protein expression, suggesting that these substances may be used in prevention and/or treatment of various human cancers like spinal, brain, colon, breast, prostate, bladder and endometrial cancers.
This was to study expression and sub-cellular localisation of PTEN protein, and review the effect(s) of indomethacin on PTEN's expression in cultured Human Endometrial Cancer (HEC 1B) cell line, which is known to express significant amounts of the wild-type PTEN.
This involves culture and incubation of artificial HEC 1B cells. All procedures were undertaken in the cell culture hood under the recommended sterile conditions. The cells were then incubated with different concentrations of indomethacin solution, for variable durations and finally fixed (with paraformaldehyde) and stained with fluorescein-labelled diluted secondary antibody (FITC). Immunocytochemistry (IHC) and fluorescent microscopy were then employed for the detection and localisation of the specific antigen (PTEN), using antibodies.
The HEC 1B cells, which were cultured and incubated with different concentrations of indomethacin solution, expressed the PTEN protein, most of which was localised to the nucleus with minimal cytoplasmic expression. Increased PTEN expression was observed following treatment of the cells with various concentrations of the solution for variable durations, although there was cell death at higher concentrations and longer duration. This procedure was repeated several times, in order to have consistency and to validate the results.
This study agrees with previous studies in similar human cell lines and supports the idea that NSAIDs and other drugs may be used in the future for prevention of human cancers. However, more studies need to be carried out to substantiate these observations.
先前的研究报道，非甾体抗炎药（NSAIDs）、化学物质和食品补充剂可用于上调PTEN mRNA和蛋白表达，这表明这些物质可用于预防和/或治疗各种人类癌症，如脊髓癌、脑癌、结肠癌、乳腺癌、前列腺癌、膀胱癌和子宫内膜癌。
本研究旨在研究PTEN蛋白的表达及亚细胞定位，并综述吲哚美辛对培养的人子宫内膜癌（HEC 1B）细胞系中PTEN表达的影响，已知该细胞系表达大量野生型PTEN。
本研究涉及人工培养的HEC 1B细胞的培养和孵育。所有操作均在细胞培养罩中按照推荐的无菌条件进行。然后将细胞与不同浓度的吲哚美辛溶液孵育不同时间，最后用多聚甲醛固定，并用荧光素标记的稀释二抗（FITC）染色。然后采用免疫细胞化学（IHC）和荧光显微镜，使用抗体检测和定位特异性抗原（PTEN）。
用不同浓度的吲哚美辛溶液培养和孵育的HEC 1B细胞表达PTEN蛋白，其中大部分定位于细胞核，细胞质表达极少。在用不同浓度的溶液处理细胞不同时间后，观察到PTEN表达增加，尽管在较高浓度和较长时间处理时有细胞死亡。为了保证一致性和验证结果，该过程重复了几次。
本研究与先前在类似人类细胞系中的研究结果一致，并支持NSAIDs和其他药物未来可能用于预防人类癌症的观点。然而，需要进行更多研究来证实这些观察结果。
