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首頁> 外文學(xué)位 >Development of an immortalized human cell line to study the effects of environmental exposure to carcinogens.
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Development of an immortalized human cell line to study the effects of environmental exposure to carcinogens.

機譯：開發(fā)永生化人類細胞系，以研究環(huán)境暴露于致癌物的影響。

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In order to provide an improved in vitro model with which to investigate human diseases, such as cancer that may be promoted by toxicant exposure, we have characterized a newly developed cell line derived from the renal proximal tubule epithelial cells (RPTEC) of a healthy human male donor. The RPTEC/TERT1 cell line has been immortalized using the human telomerase reverse transcriptase (hTERT) catalytic subunit and does not exhibit chromosomal abnormalities (Evercyte Laboratories).;We have conducted single-compound and binary mixture experiments with the common environmental carcinogens, cadmium (Cd) and benzo[a]pyrene (B[a]P). Cells exhibited cytotoxicity to concentrations of B[a]P and Cd as low as 1 nM and 3 muM, respectively. We examined a panel of eight genes relevant to the toxic responses of these two agents. RPTEC/TERT1 cells exhibit compound-specific gene expression responses to concentrations as low as 1 nM B[a]P and 1 muM Cd. A significant increase in the expression of genes coding for B[a]P metabolizing enzymes (CYP1A1, CYP1B1) occurred in a dose- and time-dependent manner. Activity of these enzymes was verified using the EROD activity assay. Gene expression changes after co-exposure were consistent with changes in gene expression seen after single-compound exposures.;We detected BPDE-DNA adducts after exposure to B[a]P which confirms that the RPTEC/TERT1 cell line responds to B[a]P consistently with what is known regarding these cells in a normal, healthy kidney. Under co-exposure, adducts detected were significantly decreased in some groups. A significant increase in the expression of NRF2 antioxidant pathway genes after co-exposure was observed. Additionally, total glutathione levels were significantly increased in cells exposed to Cd alone and co-exposure groups. These results suggest that Cd may antagonize the formation of BPDE-DNA adducts in RPTEC/TERT1 cells under these conditions.;Future studies will test mutagenesis under conditions of co-exposure to Cd and B[a]P. Our studies are the first to provide information regarding toxicological responses in this novel cell line that model those of the target tissue. We conclude that these cells can provide a useful tool for future toxicological studies. These studies will help scientists better understand the initiating events that may promote carcinogenesis in normal, healthy human cells.
機譯：為了提供改進的體外模型來研究人類疾病，例如可能通過暴露有毒物質(zhì)引起的癌癥，我們鑒定了一種新開發(fā)的，源于健康人的腎近端腎小管上皮細胞（RPTEC）的細胞系男性供體。 RPTEC / TERT1細胞系已使用人類端粒酶逆轉(zhuǎn)錄酶（hTERT）催化亞基永生化，并且沒有表現(xiàn)出染色體異常（Evercyte Laboratories）。；我們已對常見的環(huán)境致癌物鎘（ Cd）和苯并[a] py（B [a] P）。細胞對B [a] P和Cd的濃度分別低至1 nM和3μM表現(xiàn)出細胞毒性。我們檢查了與這兩種藥物的毒性反應(yīng)相關(guān)的八個基因。 RPTEC / TERT1細胞對低至1 nM B [a] P和1μMCd的濃度表現(xiàn)出化合物特異性基因表達反應(yīng)。 B [a] P代謝酶（CYP1A1，CYP1B1）編碼基因的表達顯著增加，且呈劑量和時間依賴性。使用EROD活性測定法驗證了這些酶的活性。共暴露后基因表達的變化與單化合物暴露后基因表達的變化一致。；我們在暴露于B [a] P后檢測到BPDE-DNA加合物，這證實RPTEC / TERT1細胞系對B [a]有反應(yīng)] P與正常健康腎臟中有關(guān)這些細胞的已知信息一致。在共同暴露下，某些組中檢測到的加合物明顯減少。共同暴露后，NRF2抗氧化劑途徑基因的表達明顯增加。此外，單獨暴露于鎘和共同暴露組的細胞中總谷胱甘肽水平顯著增加。這些結(jié)果表明，在這些條件下，鎘可能會拮抗RPTEC / TERT1細胞中BPDE-DNA加合物的形成。未來的研究將測試在同時暴露于鎘和B [a] P的條件下的誘變作用。我們的研究首次提供了有關(guān)這種新型細胞系中毒理學(xué)反應(yīng)的信息，該細胞系模擬了靶組織的毒理學(xué)反應(yīng)。我們得出結(jié)論，這些細胞可以為將來的毒理學(xué)研究提供有用的工具。這些研究將幫助科學(xué)家更好地了解可能促進正常健康人類細胞癌變的起始事件。

著錄項

  • 作者

    Simon, Bridget R.;

  • 作者單位

    Tulane University.;

  • 授予單位 Tulane University.;
  • 學(xué)科 Microbiology.;Toxicology.;Environmental science.;Oncology.
  • 學(xué)位 Ph.D.
  • 年度 2015
  • 頁碼 139 p.
  • 總頁數(shù) 139
  • 原文格式 PDF
  • 正文語種 eng
  • 中圖分類
  • 關(guān)鍵詞

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