Genistein及ICI182,780單獨或合併處理對肺腫瘤及 正常細胞生長的影響

The effects of genistein and ICI182,780 alone or combined on the growth of lung tumor and normal cells

陳佩樺(Pei-Hua Chen)

指導教授 ： 葉姝蘭

中山醫學大學/健康管理學院/營養學研究所/碩士(2008年)

https://doi.org/10.6834/CSMU.2008.00114

摘要

研究指出肺癌細胞estrogen receptor (ER)表現的差異，亦即ERα和/或ERβ，可能影響肺癌的發展及其對治療的反應。植物雌激素及雌激素拮抗藥物如ICI182,780 (ICI，一種抑制雌激素之類固醇化合物)均可藉由ERs調節基因而抑制乳癌細胞生長。但合併處理此兩類分子，在不同ER表現肺癌細胞中，是否影響ER表現及是否可抑制細胞生長並不清楚。因此使用兩株人類肺癌細胞株，分別為A549 (ERα/β:-/+)及NCI-H460 (ERα/β: +/+)作為研究模式，我們首先篩檢四種不同的phytoestrogens ，分別為20 μM genistein、daidzein、enterolactone及35 μM enterodiol，以及兩種抗癌藥物ICI及tamoxifen對細胞生長的影響。結果顯示，在抑制癌細胞生長作用上，phytoestrogen中以genistein最為顯著；藥物中，ICI的作用類似或較tamoxifen好。但由於20 μM genistein亦對正常肺細胞株IMR-90具生長抑制作用，因此我們以2 μM、10 μM genistein進行後續實驗，並研究2 μM、10 μM genistein及ICI單獨或合併處理後，對細胞生長的影響。結果顯示，genistein單獨處理，會抑制兩株肺癌細胞的生長，並具有劑量效應，但對IMR-90沒有毒害，genistein會使細胞週期停滯於G2/M期。而ICI則藉由使細胞停滯G0/G1期及增加sub-G1的細胞數而抑制兩株肺癌細胞生長。ICI合併2 μM genistein處理後，抑制A549細胞生長情形較ICI單獨處理時佳，但對NCI-H460細胞，此合併處理並無加強效果，在24及48小時2 μM genistein甚至降低了ICI的效果。不過，在兩株細胞中，ICI合併10 μM genistein處理後，與個別單獨處理時比較，皆增加了細胞生長停滯的效應，亦增加sub-G1期的細胞數。ERs mRNA表現方面，單獨genistein會增加兩株細胞中ERβ mRNA之表現量，並會抑制NCI-H460細胞ERα mRNA之表現；而單獨ICI處理，則抑制NCI-H460細胞ERα mRNA表現，但對兩株細胞ERβ mRNA則影響很小或沒影響。在ICI合併genistein處理後，相加性的調節ERα及ERβ mRNA的表現量。此調節ERα及ERβ mRNA表現的作用情形，似乎僅能部份解釋單獨或合併處理ICI及genistein後，對於細胞生長停滯作用之影響。總結以上結果，本實驗發現，在ICI合併genistein處理後，會因不同的肺癌細胞株而有不同的影響程度。ICI合併genistein處理後，對生長抑制作用的精確機制仍須進一步探討。

關鍵字

genistein ； ICI182,780 ； A549 ； NCI-H460 ； IMR-90 ； estrogen receptors

並列摘要

Some studies suggest that the variation of estrogen receptor (ER) expression, ERα and/or ERβ﹐in lung tumor cells may influence the responses to therapy. Phytoestrogens as well as estrogen-antagonistic drugs, such as ICI182,780 (ICI) can regulate gene expression and inhibit cell growth through ERs in breast cancer. Whether combined these compounds affect the expression of ERs and inhibit cell growth in lung cancer cells with different types of ER remain unclear. Therefore, using two human lung tumor cell lines, A549 (ERα/β: -/+) and NCI-H460 (ERα/β: +/+), we first screen the effects of 20 μM genistein, daidzein, enterolactone and 35 μM enterodiol, 4 phytoestrogens, as well as ICI and tamoxifen, 2 anticancer drugs, on the growth of A549 cells and NCI-H460 cells. The results showed that genistein had a greatest cell-growth inhibitory effect among those phytoestrogens, while the cell-growth inhibitory effect of ICI was similar to or better than that of tamoxifen. However, 20 μM genistein also inhibit the growth of human fetal lung fibroblast, IMR-90. Thus, we further investigated the individual and combined effects of genistein at 2 or 10 μM and 1 μM ICI on the cell growth of these cell lines. The results showed that genistein alone inhibited the growth of both lung cancer cell lines in a dose dependent manner, while it had no effect on the growth of IMR-90. Genistein alone induced cell cycle arrest at G2/M phase. ICI alone inhibited the growth of both lung cancer lines through inducing cell cycle arrest at G0/G1 phase and increasing sub-G1 cells. The combined effect of 2 μM genistein and ICI on cell- growth arrest was greater than the individual effects in A549 cells; whereas, at 24 and 48 h, 2 μM genistein decreased the cell-growth arrest effect of ICI in NCI-H460 cells. However, the combined treatment of 10 μM genistein and ICI enhanced cell-growth arrest effect as compared with individual in both cancer cell lines. The combination of 10 μM genistein and ICI induced an increase of sub-G1 cells (%). Genistein alone increased the mRNA level of ERβ in both cancer cell lines, while it decreased ERα mRNA expression in NCI-H460 cells. ICI alone suppressed ERα mRNA expression in NCI-H460 cells, while just had little or no effect on the expression of ERβ mRNA in both cancer cell lines. The combined treatments of genistein and ICI additively modulated the expression of ERα and ERβ mRNA. The modulation of expression of ERα and ERβ seems partly rather than completely explain the cell growth arrest effects of genistein and ICI alone or combined. In conclusion, this study demonstrated that the combined effects of genistein and ICI are different in different lung cancer lines. The precise mechanisms of the combined effects of genistein and ICI are needed more studies to investigate.

並列關鍵字

genistein ； ICI182,780 ； A549 ； NCI-H460 ； IMR-90 ； estrogen receptors

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