空間環(huán)境對惡性黑色素瘤B16細(xì)胞間通訊功能的影響

摘要： 觀察空間誘變后惡性黑色素瘤B16細(xì)胞株的細(xì)胞間通訊功能及細(xì)胞間黏附能力的變化,并探討可能的機(jī)理。應(yīng)用細(xì)胞低溫長期生存系統(tǒng),將B16細(xì)胞株搭載于“第20顆返回式衛(wèi)星”,返地后單克隆化,從得到的110株單克隆空間誘變B16細(xì)胞株中隨機(jī)選取6株,編號為39#～44#,常規(guī)培養(yǎng)6代后和對照細(xì)胞株同時(shí)做MTT和FCM,檢測細(xì)胞增殖和周期分布;用劃痕標(biāo)記染料示蹤技術(shù)檢測細(xì)胞間通訊功能;將有變異的細(xì)胞接種C57BL/6J小鼠,免疫組化觀測CD31蛋白的表達(dá)量。初步篩選出兩株(39#、44#)變化比較明顯的細(xì)胞株,44#細(xì)胞增殖能力降低、G1期分布增加、細(xì)胞間通訊功能增強(qiáng)、CD31蛋白表達(dá)增強(qiáng);39#細(xì)胞增殖能力升高、G1期分布減少、細(xì)胞間通訊功能減弱、CD31蛋白表達(dá)減少?？臻g環(huán)境的作用使體外培養(yǎng)腫瘤細(xì)胞產(chǎn)生多向變異,有可能篩選出有意義的細(xì)胞株做進(jìn)一步的研究。
To study the gap junction intercellular communication and adhension of the melanoma B16 cell clones mutagenized by space environment,and the possible mechanism.B16 cells were put on board of the Chinese 20th recoverable satellite,and were harvested after 18 days' spaceflight and then monocloned.Six cell clones numbered as 39#～44# were randomly selected from the total 110 clones obtained,and the control clones were routinely cultured.After six generations,the cultured cells were experimented with MTT assay and FCM methods for cell growth characteristics;with scrape loading dye transfer method for gap junction interceluler communication;with immunohistochemistry method for the CD31 contents.Two clones were selected as remarkably different from the control clone in growth characteristics,gap junction intercellular communication,intercellular adhension.Cultured melanoma B16 cells in vitro were found to be mutated by the outer space environment.One may screen out mutagensis B16 cell clones to carry out studies useful for human beings.