一. IL-13(介白素-13)是由第二型幫助性T淋巴球所分泌的重要細胞激素，和氣喘這種疾病有所關聯性。IL-19 (介白素-19)是屬於IL-10(介白素-10)家族成員之一而且目前對於它所扮演的生物功能了解不多。IL-21(介白素21)是第一型細胞激素家族成員之一，此外，會專一性的抑制未活化幫助性T淋巴球分化成由干擾素-r誘導的第二型幫助性T淋巴球，所以也是一種第二型幫助性T淋巴球的細胞激素。在我們之前的研究指出IL-19和IL-21和氣喘的發病機制有關，我們藉由ELISA分析氣喘病人血清中含有的IL-19和IL-21，發現氣喘病人血清中IL -19和IL-21的含量較健康個體高兩倍。有興趣去了解IL-19和IL-21作用在活化的T淋巴球對於IL-13基因表現的調控情形，我們構築兩段相對於轉錄起始點有可能為轉錄活化位置的片段在含有luciferase基因的質體上，這些片段分別包含自轉錄起始點上游的+49開始到-940及-1560的序列，簡稱為pA和pB。將這些質體transfect至Jurkat T 細胞 。Jurkat T 細胞是人類T 細胞被使用當作需要T 細胞活化的模式，當培養液加上PMA和ConA時，細胞會被刺激呈現活化狀態。藉由luciferase的活性分析，發現pA可以驅使 luciferase有較高的表現量。當IL-21不論作用在是否有活化的Jurkat T細胞，對於IL-13自轉錄起始點上游的+49開始到-940至-1560的promoter活性皆沒有影響，有可能IL-21對Jurkat T細胞IL-13基因表現是影響在mRNA的穩定性或是在其他片段上。因此，接下來我們以real-time PCR的方式去分析IL-19和IL-21作用在Jurkat T 細胞對於IL-13轉錄層面的影響，結果指出IL-19與IL-21促使活化的Jurkat T細胞表現IL-13，因為IL-19與IL-21作用於未活化的Jurkat T細胞對於IL-13的表現沒有差異，所以促使IL-13的表現，T淋巴球細胞先被活化是有必要的。

　　二. Mda-7是利用subtractive hybridization的方法從人類HO-1黑色素細胞瘤被發現的。mda-7 近來也被稱為IL-24，屬於IL-10家族成員之一。IL-24 對於廣泛性的腫瘤不論是在體內或體外都可以抑制其生長，而對正常的細胞則沒有傷害性。然而，IL-24是否具有抗肝癌的活性仍是未知的。我們的目的是以intramuscular electroporation的方式表達IL-24的基因，觀察IL-24對患有肝癌小鼠之影響，經由打入 ML-14a細胞至脾臟再轉移至肝臟而產生原位肝癌小鼠模式。結果顯示在打入ML-14a細胞至脾臟第43天，有過量表現IL-24的組別(N＝13)具有46％的存活率，相較控制組7％之存活率高，所以IL-24具有抗肝癌的活性。有過量表現IL-24的小鼠血清中IL-6的含量也較控制組高，推測IL-24能抗肝癌之機制可能和啟動免疫系統有所關聯。然而，在體外觀察IL-24對ML-14a細胞生長之情形，並沒有什麼影響。因此，有關IL-24更詳細的抗肝癌機制，有待日後進一步的探討，而在未來IL-24也許有潛力應用在治療肝癌的疾病。

　　Ⅰ. Interleukin (IL)-13 is an important cytokine secreted from type 2 helper T lymphocytes and has been associated with asthma. Interleukin (IL)-19 belongs to the IL-10 family and little was known about the biologic function. Interleukin (IL)-21 is a new member of the type I cytokine superfamily and is a T helper (Th) cell 2 cytokine that specifically inhibits the differentiation of naive Th cells into interferon gamma-producing Th1 cells. In our previous studies, we explored IL-19 & IL-21 associated with pathogenesis of asthma, we employed ELISA to analyze the serum level of IL-19 & IL-21 in the asthma patients and found that IL-19 & IL-21 level in the patients was two-fold over the normal control. To investigate the regulation of IL-13 gene expression by IL-19 & IL-21 with activation of T cell in vitro. We constructed two plasmid DNA vectors which contained different length of IL-13 promoter (-1560 to +49,-940 to +49) upstream of luciferase reporter gene. We named these two constructs, pA and pB. These two plasmids were transfected into Jurkat T cell line. Jurkat T cells were used as a model to examine the requirements of T cell activation. Cell cultures were stimulated with PMA and ConA. The results of luciferase activity showed that pA region expressed higher promoter activity. Treatment of cells with IL-21 resulted in no significant effect of luciferase activity. IL-21 may induce IL-13 transcript on the activated T cells through enhancing mRNA stability or other factors. Further, we employed real-time PCR to analyze the transcription level of IL-13 by treatment of IL-19 & IL-21 in Jurkat T cell line. IL-19 & IL-21 induced IL-13 transcript by the activated T cells. Activation of T cells is required for induction of IL-13 because IL-19 & IL-21 did not induce IL-13 production on unstimulated T cells.

　　Ⅱ. The melanoma differentiation-associated gene 7 (mda-7) was identified by a subtraction hybridization approach from the human HO-1 melanoma cell line. The mda-7 gene belongs to the IL-10 family of cytokines and has recently been classified as IL-24. Introduction of the mda-7/IL-24 gene into a wide variety of cancer cells suppressed growth in vitro and in vivo, with minimal toxicity to normal cells. However, it has not been unkown whether IL-24 has the activity of anti-hepatoma. The purpose of this study was to determine the in vivo tumor suppression function of IL-24 on the hepatoma cells by the approach of intramuscular electroporation. We generated a hepatoma mice model by injecting ML-14acells from mice spleen to allow tumor cells grown at liver. Forty-three days after injection of ML-14a cells. Forty-six percent of mice (N=13) survived from hepatoma compare to seven percent of the negative control. This result demonstrated that IL-24 is a tumor suppressor for hepatoma cells. Also, IL-6 level of mouse serum were higher than negative control. We proposed that anti-tumor mechanism of IL-24 was associated with immunostimulatory activity. However, Treatment of ML-14a cells with IL-24 resulted in no significant antiproliferative activity in vitro. The detailed mechanism of the anti-hepatoma activity of IL-24 should be further studied. IL-24 may be an effective treatment for hepatoma in the future.

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