【作者】 栗宝华；

【导师】 赵丽嫣；

【作者基本信息】 大连医科大学 ， 妇产科学， 2003， 硕士

【摘要】 卵巢癌是妇科三大恶性肿瘤之一，起病隐匿、恶性度高，完善的肿瘤细胞减灭术和术后辅以铂类为主的一线化疗是当前治疗卵巢癌的主要手段。但由于一线化疗后顺铂耐药的存在造成卵巢癌患者五年生存率较低，仅为30％左右。对铂类耐药的卵巢癌治疗是当前研究的热点。近些年来，流行病学、实验研究已证实卵巢癌是激素依赖性肿瘤，激素在卵巢癌的病因和辅助治疗中发挥重要作用；又有研究发现高浓度的雌、孕激素在体外可抑制卵巢癌耐药细胞株OVCAR-3细胞生长，并增加阿霉素对癌细胞的杀灭作用，但激素对耐药的卵巢癌细胞作用机制如何，目前国内外报道较少。三苯氧胺（tamoxifen，TAM）为雌激素受体（estrogen receptor,ER）拮抗剂，可通过与雌激素竞争受体抑制肿瘤细胞增殖；甲孕酮（medroxyprogesterone，MPA）为合成类孕激素，可通过孕激素受体（progesterone receptor，PR）抑制雌激素受体、孕激素受体阳性的卵巢癌细胞株生长。本文旨在：1．研究三苯氧胺和甲孕酮单独、联合及序贯用药对卵巢癌顺铂耐药细胞株SKOV-3/DDP生长的影响，并探讨其作用机制，以期为临床顺铂耐药患者提供新的辅助治疗方案。2．因SKOV-3/DDP细胞株存在雌激素受体变异，对其研究以探讨激素对存在受体变异的卵巢癌细胞株的作用机制。 方法 1．用四甲基偶氮唑蓝（MTT）分析法测定细胞增殖情况，2．用电子显微镜观察细胞超微结构的形态学变化，3．用流式细胞仪测定细胞周期分布及凋亡率变化情况。以比较不同浓度的TAM和MPA单独、联合及序贯用药对卵巢癌顺铂耐药细胞株SKOV-3/DDP生长的影响。 结果 1．用不同浓度(1×10^-10～1×10^-4mol/L)的TAM作用于SKOV-3/DDP细胞48小时后，低浓度(1×10^-10～1×10^-7mol/L)时细胞生长未受影响，高浓度(1×10^-6～1×10^-4mol/L)时，TAM可使G₀/G₁期细胞增加，S+G₂/M期细胞减少，而抑制肿瘤细胞增殖，且随着浓度增加，在抑制细胞增殖的同时，诱导细胞凋亡，与对照组及组间比较，差异均有显著性（P<0.01）。2.不同浓度（1 xl护0-1 xl少mo比）的MPA作用SKOV一3月ODP细胞48小时后，各浓度对细胞生长均无影响，与对照组比较，无显著性差异（P>0.05）。3.不同浓度（l x10-’0¹ x 10闷m。比）的TAM和N田A联合用药，低浓度（l x 10-10-1 xlo一b。比）时对细胞生长也无影响，高浓度（1 xlo气1 xl丫m。比）时可抑制细胞生长，且其抑制细胞生长能力较单用TAM组为强，组间比较，差异有显著性（P<0 .05）。4.1 x 10)山。比的TAM作用24h后再序贯使用各浓度的NJA，高浓度（l x 10一s一1 x 10闷m。比）时抑制肿瘤细胞生长，且其抑制细胞生长能力可被孕激素受体拮抗剂米非司酮拮抗。 结论提示: 1.TAM对卵巢癌耐药细胞株SKOV一3心DP是通过受体外机制发挥抗癌作用。 2.N[PA对卵巢癌细胞生长的影响需要功能性ER、PR的共存。 3.TAM和N田A联合及序贯用药对SKOV一3心DP生长的影响可能是通过抗UER和P用R系统而发挥作用。 4.1人M和N田A联合及序贯用药对顺铂耐药的卵巢癌细胞株生长有抑制作用，且因其较小的副作用及肯定的临床意义有可能成为卵巢癌患者化疗有效的辅助用药。更多还原

【Abstract】 Ovarian carcinoma is one of the most common fatal gynecologic malignancies in the world. It is diagnosed in an advanced state and is high in mortality. Ovarian carcinoma is current treated with cytoreductive surgery and cisplatin-basted combination chemotherapy. But patients whose tumor was resistant to cisplatin-basted chemotherapy have a poor 5-year survival and only approximate 30% 5-year survival.In recent years, the fact that ovarian carcinoma is a hormones-dependent tumor has been proved with epidemiology and experiment research. Hormones are important to etiologic factor and adjuvant therapy of ovarian tumor. The research found that estradiol and medroxyprogesterone at high concentrations could inhibit the growth of OVACAR-3 cell line and could reverse the doxorubin resistance of OVACAR-3 cell line. But which effect is it that hormones inhibit the growth of cisplatin-resistant human epithelial ovarian cancer cell line? This article is rare. Tamoxifen which is an estrogen receptor antagonist could inhibit the growth of tumor cell by competition to receptor with estrogen. Medroxyprogesterone could inhibit the growth of ovarian tumor cell including ER and PR by PR. The article’s objective are 1. to study the effects of tamoxifen and medroxyprogesterone on the growth of cisplatin-resistant human epithelial ovarian cancer cell line SKOV-3/DDP and to discuss their mechanisms. 2. to study the effects of tamoxifen and medroxyprogesterone on the growth of human epithelial ovarian cancer cell line SKOV-3/DDP involving a novel mutation of ER transcripts.Methods SKOV-3/DDP cells were treated with different concentrations tamoxifen and medroxyprogesterone. 1. The inhibition of SKOV-3/DDP cell growth was measured by methyl thiazolyl tetrazolium(MTT) rapid photocolorimetric assay. 2.The morphological change of cell was observed by electron microscopy. 3. Cell cycle and apoptotic percentage were accounted by flow cytometry.Results 1. SKOV-3/DDP cell growth was inhibited by tamoxifen at high concentration (1 × 10-6-1 × 10-4mol/L) in a dose-dependent manner (P<0.01) .Low concentration (1× 10-10~1 × 10-7mol/L) of tamoxifen had no effect on SKOV-3/DDP cell growth. With the high concentration (1×10-6-1×10-4mol/L) of tamoxifen, G0/G1 arrest of SKOV-3/DDP cell was enhanced. With the higher concentration of tamoxifen (1×10-5, 1×10-4mol/L) ,the apoptotic peak and percentage was enhanced. 2. Every concentration (1×10-10~1 ×10-4mol/L) of medroxyprogesterone had no effect on SKOV-3/DDP cell growth (P>0.05) . 3. SKOV-3/DDP cell growth was inhibited by tamoxifen combined with medroxyprogesterone at high concentration (1×10-6~1× 10-4mol/L) (P<0.01) .Low concentration (1×10-10-1X 10-7mol/L) of them had no effect on SKOV-3/DDP cell growth (P>0.05 ) . The effect of tamoxifen combined with medroxyprogesterone was more obvious than tamoxifen used separately (P<0.05 ) . 4. After SKOV-3/DDP cells were treated with tamoxifen at 1×10-7mol/L concentration in 24h, high concentration (1×10-5~1×10-4mol/L) of medroxyprogesterone inhibited the growth of SKOV-3/DDP cells. The inhibitory effect of medroxyprogesterone was significantly blocked by mifepristone which is an progesterone receptor antagonist. Conclusions1. Tamoxifen inhibits the growth of SKOV-3/DDP cells involving a novel mutation of ER transcripts by a receptor-independent mechanism.2. The inhibitory effect of medroxyprogesterone on the growth of SKOV-3/DDP cells requires association ER with PR fuctionally.3. The present study suggests that a potential interaction between the anti-E/ER and P/PR systems could be important in the growth regulation of SKOV-3/DDP cells by tamoxifen and medroxyprogesterone.4. Tamoxifen and medroxyprogesterone with the inhibitory effect on the growth of cisplatin-resistant ovarian cancer cells and the minor ill effect could be used to ovarian cancer patients as an adjuvant therapy.更多还原