本文選題：卵巢癌　切入點：雌激索　出處：《南京中醫(yī)藥大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：研究雌孕激素聯(lián)合紫杉醇對人卵巢癌細(xì)胞體外生長的調(diào)控作用,并且探索改變細(xì)胞生長狀態(tài)的可能機(jī)制,為其在臨床中用于卵巢癌的輔助治療提供前期的理論依據(jù)。 方法：梯度濃度紫杉醇體外作用人卵巢癌細(xì)胞,四甲基偶氮唑蘭(methyl thiazolyl tetrazolium, MTT)比色法檢測藥物作用24h、48h、72h后細(xì)胞的抑制率；根據(jù)IC50確定下一階段實驗藥物的濃度。根據(jù)前期實驗結(jié)果,10－4nol/L雌孕激素抑制卵巢癌細(xì)胞生長。組別設(shè)置為對照組(control)、雌激素組(E2)、孕激素組(P4)、紫杉醇組(Taxol)、雌激素聯(lián)合紫杉醇組(E2+Taxol)、孕激素聯(lián)合紫杉醇組(P4+Taxol),各組藥物作用于卵巢癌細(xì)胞48h。 MTT檢測各組卵巢癌細(xì)胞的抑制率；流式細(xì)胞儀檢測各組卵巢癌細(xì)胞的凋亡率和細(xì)胞周期；實時熒光定量PCR (qRT-PCR)法和蛋白質(zhì)印跡(Western blot)法檢測各組卵巢癌細(xì)胞drosha基因和蛋白的表達(dá)。 結(jié)果：①MTT顯示：E2(10-4mol/L)、P4(10-4mol/L)、Taxol (10ug/ml)、E2+Taxool、 P4+Taxol均能抑制人卵巢癌細(xì)胞生長,誘導(dǎo)細(xì)胞凋亡：雌激素對ER(-)的卵巢癌細(xì)胞生長抑制作用強(qiáng)于ER(+)的卵巢癌細(xì)胞(P0.05),并且與紫杉醇具有協(xié)同作用；孕激索對卵巢癌細(xì)胞也具有生長抑制作用(P0.05)。②流式細(xì)胞技術(shù)檢測細(xì)胞凋亡顯示：紫杉醇可以提高卵巢癌細(xì)胞的凋亡率,與對照組比較具有顯著性差異(P0.05)：雌激素作用卵巢癌細(xì)胞后凋亡率增加,且.ER(-)的卵巢癌細(xì)胞凋亡率高于ER(+)的卵巢癌細(xì)胞,雌激素聯(lián)合紫杉醇凋亡率也增加；孕激素單用或聯(lián)合紫杉醇,卵巢癌細(xì)胞凋亡率也有所提高(P0.001)。③流式細(xì)胞技術(shù)檢測細(xì)胞周期顯示：雌激素作用后大量細(xì)胞阻滯在S期,孕激素作用后細(xì)胞周期無明顯改變,紫杉醇作用后大量細(xì)胞阻滯在G2/M期。④qRT-PCR和Western blot結(jié)果顯示：紫杉醇能上調(diào)drosha基因及蛋白表達(dá)(P0.001)：雌激素對drosha基因及蛋白的影響與卵巢癌細(xì)胞ER表達(dá)相關(guān)：ER(-)者,drosha基因及蛋白表達(dá)上調(diào)(P0.001)：ER(+)者,drosha基因及蛋白表達(dá)差異無統(tǒng)計學(xué)意義。兩藥聯(lián)合作用后,drosha基因及蛋白的表達(dá)較對照組上調(diào)(P0.001)；孕激素、孕激素聯(lián)合紫杉醇均能上調(diào)drosha基因及蛋白表達(dá)(P0.001),但兩藥聯(lián)合較紫杉醇組差異無統(tǒng)計學(xué)意義。 結(jié)論：雌、孕激素聯(lián)合紫杉醇能夠抑制體外培養(yǎng)的人卵巢癌細(xì)胞的生長,這三種藥物均能改變細(xì)胞的凋亡率,雌激素和紫杉醇能夠改變細(xì)胞周期。雌孕激素聯(lián)合紫杉醇通過上調(diào)drosha表達(dá),起到抑癌或者增敏作用,并且與雌激素受體的表達(dá)相關(guān)。
[Abstract]:Objective: to study the effects of estrogen and paclitaxel on the growth of human ovarian cancer cells in vitro, and to explore the possible mechanism of changing the growth state of human ovarian cancer cells, and to provide a theoretical basis for the clinical application of estrogen and paclitaxel in the adjuvant treatment of ovarian cancer. Methods: human ovarian cancer cells were treated with gradient concentration of paclitaxel in vitro. The inhibition rate of the cells was determined by MTT colorimetry after 24 h, 48 h and 72 h exposure to methylazolam methyl tetrazolium (MTT). Determine the concentration of the next phase of the experimental drug based on IC50. According to the previous experimental results, 10-4 nol-1 / L estradiol inhibits the growth of ovarian cancer cells. The group is divided into three groups: control group, estrogen group, estradiol group, progesterone group, taxolol group, estrogen combined with purple. Estradiol group, progesterone combined with paclitaxel group, P4 Taxolol group. The inhibition rate of ovarian cancer cells in each group was detected by MTT for 48 h. The apoptosis rate and cell cycle were detected by flow cytometry, and the expression of drosha gene and protein were detected by real-time quantitative PCR qRT-PCR and Western blotting. Results: the results showed that 10 ~ (-4) mol / L ~ (-1) Taxol of E _ (2) N _ (10 ~ (-4)) P _ (10 ~ (-4)) P _ (4) Taxol could inhibit the growth of human ovarian cancer cells and induce apoptosis: estrogen had a stronger inhibitory effect on the growth of human ovarian cancer cells than that on ER () ovarian cancer cells, and had synergistic effect with paclitaxel. The results of flow cytometry showed that paclitaxel could increase the apoptosis rate of ovarian cancer cells. Compared with the control group, there was a significant difference (P 0.05): the apoptosis rate of ovarian cancer cells treated with estrogen was increased, and the apoptosis rate of ovarian cancer cells with. ER-) was higher than that of ovarian cancer cells with ER- (). The apoptosis rate of estrogen combined with paclitaxel was also increased. The apoptosis rate of ovarian cancer cells was also increased by progesterone alone or in combination with paclitaxel. The results of flow cytometry showed that a large number of cells were blocked in S phase after estrogen treatment, but the cell cycle did not change after progesterone treatment. After paclitaxel treatment, a large number of cells were blocked in G _ 2 / M phase .4qRT-PCR and Western blot. The results showed that paclitaxel could up-regulate the expression of drosha gene and protein (P0.001). The effect of estrogen on drosha gene and protein was related to ER expression of ovarian cancer cells. There was no significant difference in the expression of drosha gene and protein between the two groups. The expression of drosha gene and protein was higher than that of the control group. Progesterone combined with paclitaxel could up-regulate the expression of drosha gene and protein, but there was no significant difference between the two groups. Conclusion: estrogen and progesterone combined with paclitaxel can inhibit the growth of human ovarian cancer cells in vitro. Estrogen and paclitaxel can change cell cycle. Estrogen and progesterone combined with paclitaxel can inhibit cancer or increase sensitivity by upregulating the expression of drosha and is related to the expression of estrogen receptor.
【學(xué)位授予單位】：南京中醫(yī)藥大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R737.31

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