| Objective: Thyroid cancer is not only the highest incidence rate of malignant tumordiseases of the endocrine system, but also the most common malignant tumor of thyroid, accounting for about 1% of all malignant tumors. The papillary thyroid carcinoma(papillary thyroid carcinoma, PTC) is the most commonPathological type[1], accounting for about 60% of adultthyroid cancer and all of thyroid cancer in childhood. In recent decades, the incidence rate of thyroid papillary carcinomashowing a rapid growth trend in the world. However, the degree of malignancy of thyroid papillary carcinoma is low, even if the earlier emergence of cervical lymph node metastasis, but the prognosis is good. The 10 year survival rate could remain above 90%. At present a comprehensive treatment of papillary thyroid carcinoma recognized standard is auxiliary treatment operation resection and postoperative. Surgical treatment is the basic method of treatment of papillary thyroid cancer, adjuvant therapy may further reduce the recurrence rate, Endocrine therapy plays an important role inpostoperative adjuvant. Endocrine therapy is that the Papillary thyroid cancer patients after surgery should take thyroxine tablets for life. Through hypothalamic- pituitary- thyroid axis of self-regulation to complete. Its significance lies in the prevention of hypothyroidism and thyroid stimulating hormone(thyroid stimulating inhibited hormone, TSH). The main basis of endocrine treatment of papillary thyroid carcinoma is the presence of TSH receptor expression in papillary thyroid carcinomacells, and with the increased degree of tumor differentiation, the TSH receptor expressed the higher. With oral drug intake of thyroxine, human body endocrine TSH is reduced, thereby reducing the TSH of residualcancer tissue stimulation.Most clinical scholars believe that long-term use of thyroid stimulating hormone of patients with papillary thyroid carcinoma(TSH), inhibitory dose of thyroxine can reduce the recurrence rate and the mortality rate [2, 3]. Especially in the high-risk group of papillary thyroid carcinoma, Endocrine therapy can improve disease-free survival of patients 2-3 times[4].However, long-term use in patients with thyroid will have a certain amount of super-physiological side effects. Long term inhibition of TSH patients would lead to accelerated heart rate,frequent premature atrial contraction. According to statistics, the inhibition of TSH to the incidence of AFincreased 3 times[5] than normal. And long-term inhibition of TSH can lead to the risk of osteoporosis in postmenopausal women increased, but this does not generally occur in premenopausal women[6]. And TSH must play the role to the thyroid stimulating hormone receptorwith thyroid cell membrane or cytoplasm(TSHR) combined. Research has shown that: there is a missing of TSHR gene expression in thyroid cancer [7]. The positive expression of TSHR in papillary thyroid carcinoma rate is low, and with the clinical differentiated thyroid cancer stages increasedthe positive expression rates of TSHR declined gradually [8]. In papillary thyroid carcinoma,high levels of TSH and low expression of TSH formed the contradiction. The reason for this is not very clear, but it is certain that: there are a variety of mechanisms in complex action between thyroid stimulating hormone and thyrotropin receptor action [9].In recent years, the academic community is still controversial in whether the inhibition of thyroid papillary carcinoma is effective or not for TSH treatment. we found in clinical practice: a part of patients with papillary thyroid cancer after operation, still showed tumor recurrence and lymph node metastasis after a number of years with regular oral thyroxine preparation treatment, who need the secondary surgery. Therefore, in order to effectively reduce TSH suppressive therapy withoccurrence of side, There is important significance to the implementation of individualized treatment. The purpose of this study was to investigate the role and significance ofcomprehensive cultivation of TSH in papillary thyroid carcinoma cell in vitro.Methods: There are 5 cases confirmed to be thyroid carcinoma tissues in thyroid surgery by intraoperative frozen and paraffin after pathologically papillary,which taken from The Second Hospital of Hebei Medical University, thyroid surgery between December 2014 and January 2015. All papillary thyroid cancer patients without discharge, chemotherapy.Take the fresh tissue planted in flasks in 30 minutes, culturing papillary thyroid carcinoma cells.To be grown into single cells, we would divide into four groups and treate with different concentrations of TSH(0m U/L, 10 m U/L, 50 m U/L, 100 m U/L).And then culture in 24-well plates, detecting the proliferation and apoptosis of the cells by flow cytometry in the first 10 days of experiment.The data using t test, inspection standard for P < 0.05 had statistical significance.Result: 1 The cultivation and observation of thyroid papillary carcinoma cellsPapillary thyroid cancer cells grown in culture flask directly,the cells growth is slow.After 24 h cell culture,A few cells attached to the bottom of the culture bottle.The morphology of the cells showed round and spindle.Cells extend pseudopodia.the cells can not move any more.Cells were mainly started from the group of cells adherent.But a large number of cell suspension was not attached.These cells appeared round, moving with the culture medium.After 4~5 days, a large number of cells became adherent.Enter the incubation period of papillary thyroid cancer cells,Enter the incubation period of papillary thyroid cancer cells.5-7 days later, the cells into growth and rapid proliferation cells continue to stretch and to grow, and connected to each other,the long fusiform, almost no circular and elliptic cells, has been covered with the bottom of the culture bottle above average.After 7-8 days, the cells formed a monolayer cells in the culture bottle,and the round nucleolus and other organelles can be seen with high power microscope. 2 Comparison between the cells with different concentration of TSH culture 2.1 Screening of proliferation results from flow cytometry cell cycle, calculating multiple samples t test by SPSS 13.0 software.P < 0.05, suggesting that between the groups was statistically significant,and drawing histogram.Measurement of Pearson Correlation R2=0.955, shows good correlation and were positively correlated. 2.2 According to the results of the determination of cell apoptosis by flow cytometry, are less than 5%.Conclusion:1 TSH could induce the proliferation of in vitro cultured thyroid papillary carcinoma cells, and with the increase of TSH concentration, the ability of inducing proliferation become stronger.2 TSH has no effect on inducing apoptosis for Papillary thyroid carcinoma cells in vitro.3 The primary culture of technology of papillary thyroid carcinoma cells is Suit for studying the functional aspects of papillary thyroid cancer. It not only fill the research gap in living cells function which Other research methods can not,but also provides a more suitable alternative tools For the development of the mechanism and characteristics of papillary thyroid cancer occurrence. |
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