| | | 卵巢癌患者exosomes来源初探
| | 免费小论文 作者：李奇灵 孙阳珍 于月成 方静 张媛丽 辛晓燕
【关键词】 exosomes；腹水；卵巢肿瘤；细胞培养技术
Comparison of extraction methods of exosomes from patients with ovarian cancer
【Abstract】 AIM: To compare the extraction methods of exosomes from tissue or ascites of an advanced ovarian cancer patient, to detect the expression of exosomes of two source, and to explore the best pathway of acquiring exosomes. METHODS: The tissue of ovarian cancer was taken and the cancer cells were separated and cultured. The cell culture supernatant was collected. The ascites was collected from the same patient. The exosomes were extracted by ultracentrifugation and ultrafiltration from the cell culture supernatant and the ascites and were examined under electron microscope. HSP70, HSP90, MHC class Ⅰ molecule and MHC class Ⅱ molecule on exosomes were tested by Western blotting. The protein concentration of exosomes was tested by BCA method. RESULTS: The time for the centrifugation of ascites was longer than that of supernatant. The exosomes could be obtained from all of the supernatants and the ascites with cancer cells, not from the ascites without cancer cells. The exosomes expressed MHC class Ⅰ molecule, HSP70 and HSP90 except MHC class Ⅱ molecule. The protein concentration of exosomes in the ascites was higher than that of the supernantant of cell culture (P<0.05). CONCLUSION: Ascites is a better source of exosomes for the ovarian cancer patient with malignant effusion with positive cancer cells. The exosomes can be extracted from ovarian cancer tissue for the patients without ascites or with ascites in which cancer cells are negative.
【Keywords】exosomes; ascites; ovarian neoplasms; cell culture techniques
【摘要】目的免费小论文： 同一晚期卵巢癌患者，对组织来源和腹水来源的免费小论文exosomes进行方法和数量的比较，探讨获得卵巢癌患者exosomes的最佳途径. 方法： 收集晚期卵巢癌患者卵巢癌组织，分离癌细胞进行培养，留上清液，取同一患者的腹水，通过超速离心和超滤等方法提取上清液及腹水中的exosomes,电镜下检测；Western Blot测定exosomes 上HSP70, HSP90, MHCⅠ类分子和MHCⅡ类分子的表达；用BCA蛋白测定法测定exosomes的蛋白浓度,计算蛋白总含量. 结果： 腹水离心时间比上清液离心时间长，所有细胞培养上清液和癌细胞阳性的腹水均能分离出exosomes，癌细胞阴性的腹水中未分离出exosomes；两种来源的exosomes共同表达MHCⅠ类分子、HSP70和HSP90，均不表达MHCⅡ类分子，腹水中分离出来的exosomes蛋白含量比细胞培养上清液的含量高（P<0.05）. 结论： 腹水中癌细胞阳性的患者，腹水是exosomes的比较理想的来源；无腹水或腹水中癌细胞阴性的患者，exosomes可来源于卵巢癌组织.
【关键词】 exosomes；腹水；卵巢肿瘤；细胞培养技术
0引言
exosomes是许多种细胞都能分泌的直径30～90 nm的小囊泡，由蛋白质和脂质组成的亚细胞器. 近年来，癌症患者的肿瘤细胞和树突状细胞（dendritic cell, DC）分泌的exosomes，由于它携带所分泌细胞的一些抗原信息，被作为肿瘤特异性抗原的来源，用于恶性肿瘤免疫治疗［1-2］的研究. 由DC分泌的exosomes已经在黑色素瘤和非小细胞肺癌进入了临床Ⅰ期试验［3］, 但exosomes在卵巢癌中的研究进展较慢. 本研究我们对晚期卵巢癌患者组织来源和腹水来源的exosomes分离方法、获得量比较，探讨卵巢癌患者最佳exosomes来源途径.
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