Lunasin對肥胖因子影響MCF-7與MDA-MB-231人類乳癌細胞生長之探討
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2018
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流行病學研究顯示，過重或肥胖會增加女性罹患乳癌的風險，而肥胖過程中脂肪組織會有巨噬細胞浸潤情形，伴隨著促發炎性細胞激素的分泌而營造出一個慢性發炎的環境，適合腫瘤發展。此外，過多脂肪累積會增加aromatase的表現與活性，導致雌二醇生成增加，促使乳癌惡化。Lunasin為一種子胜肽，由43個胺基酸組成，目前研究證實具有抗腫瘤、抗發炎、抗氧化…等生理功能。然而Lunasin對肥胖相關人類乳癌細胞中的發炎介質、雌激素及芳香酶表現仍然未知。本實驗目的為探討Lunasin的介入是否會抑制肥胖微環境下雌激素依賴型MCF-7細胞及非依賴型MDA-MB-231乳癌細胞的生長並抑制其發炎反應及芳香酶表現。第一部分探討肥胖微環境是否影響乳癌發展。結果顯示在肥胖微環境下，以DMBA誘發癌化之NIH/3T3其foci數量會增加。在動物實驗發現，以DMBA誘發肥胖小鼠的乳癌腫瘤中，其腫瘤中細胞增殖能力(Ki67)與巨噬細胞浸潤(F4/80)的表現增加，副睪脂肪的M1型巨噬細胞(CD11c)及F4/80的表現也會上升，高脂飲食會降低M2型巨噬細胞(CD206)的表現，而Lunasin介入可抑制高脂飲食中的F4/80及CD11c的表現。第二部分為探討Lunasin對兩株雌激素依賴型相異的人類乳癌細胞之影響。結果顯示，MCF-7細胞在48小時之IC50為92.5 μM而MDA-MB-231細胞為136.3 μM。將細胞培養在3T3脂肪細胞條件培養液(Adipocyte conditioned medium, Ad-CM)環境下，Lunasin的處理皆能顯著抑制其兩株細胞的存活率。在細胞週期的部分，肥胖微環境下，Lunasin有將MCF-7細胞停滯在G2/M期的能力。細胞活性分析中，無論在有無Ad-CM的環境下， Lunasin的介入皆會抑制MCF-7的細胞活性，而MDA-MB-231僅在新鮮培養基下降低。另外，兩株細胞在新鮮培養基下經Lunasin處理後細胞凋亡皆增加，且芳香酶活性皆下降。
綜上所述，Lunasin可降低肥胖小鼠巨噬細胞的發炎反應，抑制乳癌細胞生長，並可降低乳癌細胞活性，增加細胞凋亡，並抑制芳香酶活性。因此，推測出富含Lunasin的飲食型態可能有助於乳癌患者的輔助治療，尤其是雌激素依賴型的乳癌。
The epidemiological studies have showed that the women with overweight or obesity have higher risk developed breast cancer. In adipogenesis, macrophages infiltrate into adipose tissue and accompany pro-inflammatory mediator secretions promoting cancer progression. In addition, excecive fat accumulation increase the expression and activity of aromatase, causing estradiol overproduction and promote the progression of breast cancer. Lunasin is a soy pepride which consist of 43 amino acids, the bioactivities of lunasin on chemopreventation, anti-inflammation and anti-oxidation have been widely studied. However, the effect of lunasin on obesity-related inflammation mediators secretion, estrogen production and aromatase expression in breast cancer cells are still uncertained. The aim of this work is to investigate whether lunasin can inhibit the growth of MCF-7 and MDA-MB-231 cells, and downregulate the inflammation in the microenvironment. First part is to find out the effect of obesity-related microenvironment on the progression of breast cancer. The results showed that that the number of DMBA-induced foci formation upregulated in obesity microenvironment in vitro studied. In vivo, F4/80, a macrophages infiltration marker, and cell proliferation increased in DMBA-induced mammary tumor in obese mice. Also, F4/80 and CD11c, a M1 macrophages marker, increased in Epididymal adipose tissue when DMBA intervention. Additionally, CD206, a M2 macrophages marker, decreased in obese mice. In addition, lunasin inhibit F4/80 and CD11c in obese mice. Second part is to investigate the effect of lunasin on estrogen dependent and estrogen independent breast cancer cells. The result revealed that after treatment with lunasin for 48h, the IC50 values in MCF-7 cells is 92.50 μM and in MDA-MB-231 cell is 136.3 μM. When incubate in 3T3-L1 adipocyte conditioned medium(Ad-CM), lunasin can inhibit cell viability in both MCF-7 and MDA-MB-231, and cell cycle arrested at G2/M phase in MCF-7. In cell vitality assay, no matter incubation with Ad-CM or not, lunasin intervention increase low vitality cells in MCF-7, but this result only found in MDA-MB-231 when incubation with fresh medium. At last, lunasin intervention increase apoptosis and decrease aromatase activity in both of breast cancer cells. In summary, lunasin downregulate inflammation in obese mice, inhibit the growth, vitality and aromatase activity and upregulate the apoptosis of breast cancer cells, which indicate that lunasin-enrich dietary life syle might be a promising adjuvant therapy to obesity-related cancer, especially estrogent dependent breast cancer.
The epidemiological studies have showed that the women with overweight or obesity have higher risk developed breast cancer. In adipogenesis, macrophages infiltrate into adipose tissue and accompany pro-inflammatory mediator secretions promoting cancer progression. In addition, excecive fat accumulation increase the expression and activity of aromatase, causing estradiol overproduction and promote the progression of breast cancer. Lunasin is a soy pepride which consist of 43 amino acids, the bioactivities of lunasin on chemopreventation, anti-inflammation and anti-oxidation have been widely studied. However, the effect of lunasin on obesity-related inflammation mediators secretion, estrogen production and aromatase expression in breast cancer cells are still uncertained. The aim of this work is to investigate whether lunasin can inhibit the growth of MCF-7 and MDA-MB-231 cells, and downregulate the inflammation in the microenvironment. First part is to find out the effect of obesity-related microenvironment on the progression of breast cancer. The results showed that that the number of DMBA-induced foci formation upregulated in obesity microenvironment in vitro studied. In vivo, F4/80, a macrophages infiltration marker, and cell proliferation increased in DMBA-induced mammary tumor in obese mice. Also, F4/80 and CD11c, a M1 macrophages marker, increased in Epididymal adipose tissue when DMBA intervention. Additionally, CD206, a M2 macrophages marker, decreased in obese mice. In addition, lunasin inhibit F4/80 and CD11c in obese mice. Second part is to investigate the effect of lunasin on estrogen dependent and estrogen independent breast cancer cells. The result revealed that after treatment with lunasin for 48h, the IC50 values in MCF-7 cells is 92.50 μM and in MDA-MB-231 cell is 136.3 μM. When incubate in 3T3-L1 adipocyte conditioned medium(Ad-CM), lunasin can inhibit cell viability in both MCF-7 and MDA-MB-231, and cell cycle arrested at G2/M phase in MCF-7. In cell vitality assay, no matter incubation with Ad-CM or not, lunasin intervention increase low vitality cells in MCF-7, but this result only found in MDA-MB-231 when incubation with fresh medium. At last, lunasin intervention increase apoptosis and decrease aromatase activity in both of breast cancer cells. In summary, lunasin downregulate inflammation in obese mice, inhibit the growth, vitality and aromatase activity and upregulate the apoptosis of breast cancer cells, which indicate that lunasin-enrich dietary life syle might be a promising adjuvant therapy to obesity-related cancer, especially estrogent dependent breast cancer.
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Keywords
Lunasin, 肥胖, 乳癌, 巨噬細胞, Lunasin, Obesity, Breast Cancer, Macrophage