Ovarian cancer cell invasiveness is associated with discordant exosomal sequestration of Let-7 miRNA and miR-200.
Ovarian cancer cell invasiveness is associated with discordant exosomal sequestration of Let-7 miRNA and miR-200.
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DOI：
10.1186/1479-5876-12-4
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发表时间：
2014-01-06
影响因子：
7.4
通讯作者：
Rice GE
中科院分区：
文献类型：
--
作者：
Kobayashi M;Salomon C;Tapia J;Illanes SE;Mitchell MD;Rice GE
外切体在癌症的发病机制和转移扩散中的作用仍有待充分阐明。最近的研究支持这一假设，即从细胞中释放外切体可以改变局部细胞外条件，促进细胞生长和新生血管形成。此外，外切体还可以改变亲本和/或靶细胞的表型。例如，将信号介体隔离到胞外体可能会降低它们在细胞内对亲本细胞的生物利用度，从而改变细胞的表型和转移潜力。释放的外切体与靶细胞的融合和传递也可能改变细胞的功能和活性。为了进一步阐明外切体在卵巢癌中的作用，本研究比较了两种不同侵袭能力的卵巢癌细胞系外切体的释放量和它们的miRNA含量。需要检验的假设是，卵巢癌细胞的侵袭性与外切体的改变释放和miRNA的外切体不一致的隔离有关。用高侵袭性(SKOV-3)和低侵袭性(OVCAR-3)卵巢癌细胞株表征其外切体的释放。SKOV-3和OVCAR-3细胞(DMEM，20%无外显子胎牛血清)在8%O2气氛中培养24小时。收集细胞条件培养液，用差速密度离心法和浮力密度离心法分离外切小体，并用Western印迹(CD63和CD9)进行鉴定。用实时荧光定量聚合酶链式反应(Real-time PCR)建立外体microRNA(let-7a-f和miR-200a-c)含量。通过典型杯状球形小泡的存在和外切体标志物CD63、CD9的表达来鉴定外切体。SKOV-3细胞释放的外切体(1.22 ± 0.11μg/106cell)是OVCAR-3(0.44 ± 0.05μg/106细胞)的2.7倍。在卵巢癌细胞系及其外体中均发现了let-7家族miRNA转录本。LET-7家族转录产物在OVCAR-3细胞中的含量高于SKOV-3细胞。相反，在SKOV-3的外体中，let-7家族的转录本比OVCAR-3中的更丰富。MIR-200家族的转录本仅在OVCAR-3细胞及其外体中发现。这项研究中获得的数据与以下假设相一致，即不同卵巢癌细胞株之间外切体的释放明显不同，并与它们的侵袭潜力有关。
The role of exosomes in the pathogenesis and metastatic spread of cancer remains to be fully elucidated. Recent studies support the hypothesis that the release of exosomes from cells modifies local extracellular conditions to promote cell growth and neovascularisation. In addition, exosomes may modify the phenotype of parent and/or target cell. For example, sequestration of signaling mediators into exosomes may reduce their intracellular bioavailability to the parent cell thereby altering cell phenotype and metastatic potential. The fusion of released exosomes with target cell and delivery may also modify cell function and activity. In this study, to further elucidate the role of exosomes in ovarian cancer, the release of exosomes from two ovarian cancer cell lines of different invasive capacity and their miRNA content of exosomes were compared. The hypothesis to be tested was that ovarian cancer cell invasiveness is associated with altered release of exosomes and discordant exosomal sequestration of miRNA. High (SKOV-3) and low (OVCAR-3) invasive ovarian cancer cell lines were used to characterize their exosome release. SKOV-3 and OVCAR-3 cells were cultured (DMEM, 20% exosome-free FBS) under an atmosphere of 8% O2 for 24 hours. Cell-conditioned media were collected and exosomes were isolated by differential and buoyant density centrifugation and characterised by Western blot (CD63 and CD9). Exosomal microRNA (let-7a-f and miR-200a-c) content was established by real-time PCR. Exosomes were identified with by the presence of typical cup-shaped spherical vesicle and the expression of exosome markers: CD63, CD9. SKOV-3 cells released 2.7-fold more exosomes (1.22 ± 0.11 μg/106 cells) compared to OVCAR-3 (0.44 ± 0.05 μg/106 cells). The let-7 family miRNA transcripts were identified in both ovarian cancer cell lines and their exosomes. The let-7 family transcripts were more abundant in OVCAR-3 cell than SKOV-3 cells. In contrast, let-7 family transcripts were more abundant in exosomes from SKOV-3 than OVCAR-3. miR-200 family transcripts were only identified in OVCAR-3 cells and their exosomes. The data obtained in this study are consistent with the hypothesis that the releases of exosomes varies significantly between ovarian cancer cell lines and correlates with their invasive potential.
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影响因子：
254.7
作者：
Jemal, Ahmedin;Siegel, Rebecca;Thun, Michael J.
通讯作者：
Thun, Michael J.
影响因子：
24.5
作者：
Paranjape T;Slack FJ;Weidhaas JB
通讯作者：
Weidhaas JB
影响因子：
3.7
作者：
Hedlund M;Nagaeva O;Kargl D;Baranov V;Mincheva-Nilsson L
通讯作者：
Mincheva-Nilsson L
影响因子：
19.6
作者：
Alvarez, M. Lucrecia;Khosroheidari, Mahdieh;DiStefano, Johanna K.
通讯作者：
DiStefano, Johanna K.
影响因子：
8.8
作者：
Taylor, DD;Gerçel-Taylor, C
通讯作者：
Gerçel-Taylor, C