| Tongue is the main position of oral cancer. Recent years, the incidence of this disease is increasing higher, and especially female patients. The prognosis of female tongue patients is worse than man's. The reason which more female patients get tongue cancer is not clear, but it suggests that endocrine factor maybe have some extent effects on the occurrence and development of tumor.It is well known that hormone and hormone-like substances are potent factors in controlling cell division and proliferation. It has been revealed that estrogen receptor (ER) exists not only in many kinds of target organ and tissue, such as uterus and mammary tissue, but also in many not traditional target tissue where people can examine the existence of estrogen receptors in differentextent, for example liver, kidney, colon, oral mucoma and salivary gland, even skin. Later people have found that the estrogen receptor also exist in many tumors, such as reproduction system tumor, stomach cancer, colon and rectal cancer, pancreatic cancer, esophageal squamous carcinoma, lung squamous carcinoma, skin tumor, and so on. The distribution of ER in tumor proposes that estrogen could control tumor occurrence, development, growth and proliferation through estrogen receptors. It is unknown whether estrogen receptors are expressed hi human tongue squamous carcinoma cell and estrogen can regulate the proliferation and metabolism. To study thoroughly on this regulation effect will help us know the mechanism of occurrence and development of human tongue squamous carcinoma.In our experiments, we firstly used methods of immunocytochemistry and RT-PCR to observe the expression of estrogen receptors in human tongue squamous carcinoma. Secondly, MTT and 3H-TdR incorporation assay were employed to investigate the influence of p-estradiol( P-E2) on the proliferation and metabolism of human tongue squamous cancer cells. Thirdly, we used flow cytometery (FCM) to study the effect of P-E2 on the cell cycle of human tongue squamous cancer cells. And finally, laser scanning confocal Microscope were employed to assay the influence of cytoplasm free calcium level ([Ca2+]) of human tongue squamous cancer cells under P-E2.The main results were as follows:1. The result of immunocytochemistry shows that there are estrogen receptors immunoreactive substances located in the human tongue squamous carcinoma. Among these, estrogen receptor-alpha (ERct) stains more strongly and this immunoreactive substances are mainly localized in the cell nucleus. However, the staining of estrogen receptor-beta (ERP) is weaker than that of ERa and its immunoreactive substances are localized in the cytoplast.2. The results of RT-PCR indicate that there are expression of ER mRNA in human tongue squamous cancer cells, including ERa and ERp. Similarly,the expression of ERp is weaker than that of ERa. Together with the data of immunoreactive substances of ER existing in these cells, it indicates that ER is expressed in human tongue squamous cancer cells and estrogen may have an influence on the proliferation and metabolism of human tongue squamous cancer cells through ER.3. The results of MTT and 3H-TdR incorporation assay showed that p-estradiol could promote the proliferation and DNA synthesis of human tongue squamous cancer cells in a dose-dependent manner (P<0.01). This effect could be block by Tamoxifen, and antogonist of estrogen receptor, which suggests that estrogen could stimulate the proliferation and metabolism of human tongue squamous cancer cells through ER.4. Flow cytometery (FCM) assay showed that, after 24h cultured with 10"6 mol/L P-E2, the proportion of GI phase decreased from (76.5 + 2.55) % to(62.3 + 3.1) %, the proportion of S phase and 62 phase increased from (23.5 + 3.52) % to (37.7+4.17) . These results indicated that 0-E2 (10"6 mol/L) could increase the proportion of cells in s and 62 phase, but reduce the proportion of cells in GI phase (PO.Ol). The cell proportion of different phase was not differen...
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