蒜素抑制人甲狀腺乳頭狀癌TPC-1細胞生長的作用及機制

翟健 韓曉晨 閆金銀

[摘要] 目的 探討蒜素（Allicin）抑制人甲狀腺乳頭狀癌TPC-1細胞生長的作用及機制。方法 將培養的TPC-1細胞分為對照組和不同濃度（10、20、30、40、50 μg/mL）Allicin處理組。四甲基偶氮噻唑藍（MTT）法測定不同濃度的Allicin處理24、48、72 h對TPC-1細胞生長的抑制作用。倒置顯微鏡和掃描電鏡觀察Allicin處理前后細胞形態和超微結構的變化。流式細胞術檢測Allicin對細胞周期的影響。免疫細胞化學技術檢測Allicin作用細胞后凋亡蛋白Bax和抗凋亡蛋白Bcl-2表達的變化。 結果 與對照組比較，不同濃度的Allicin均可抑制TPC-1細胞的生長，差異有統計學意義（P < 0.05）;Allicin對細胞的抑制率與時間和濃度兩大要素相關。隨著Allicin濃度的增加，G2/M期細胞所占比例明顯升高，G0/G1期細胞所占比例降低，除對照組與10 μg/mL Allicin組比較差異無統計學意義（P > 0.05），其他相鄰濃度組兩兩比較差異均有統計學意義（P < 0.05）。與對照組比較，各濃度Allicin處理組細胞內的Bcl-2表達量均降低，Bax表達量均升高，差異有統計學意義（P < 0.05）;各相鄰濃度Allicin處理組Bcl-2及Bax表達量比較，差異均有統計學意義（P < 0.05）。 結論 Allicin具有抑制甲狀腺乳頭狀癌TPC-1細胞生長的作用，其機制可能與Allicin抑制細胞增殖，下調Bcl-2表達，上調Bax表達從而誘導細胞凋亡有關。

[關鍵詞] 蒜素;甲狀腺乳頭狀癌;細胞周期;Bcl-2;Bax

[中圖分類號] R736.1? ? ? ? ? [文獻標識碼] A? ? ? ? ? [文章編號] 1673-7210（2019）02（a）-0013-05

[Abstract] Objective To investigate the effect and mechanism of Allicin inhibition on the growth of human thyroid papillary carcinoma TPC-1 cells. Methods The cultured TPC-1 cells were divided into the control group and the Allicin treatment groups with different concentrations （10， 20， 30， 40， 50 μg/mL）. Methylthiazoletrazolium （MTT） method was used to determine the inhibitory effect of different concentrations of Allicin treatment on the growth of TPC-1 cells at 24， 48， 72 h. Inverted microscope and scanning electron microscope were used to observe the change of cell morphology and ultrastructure after treatment by Allicin. Flow cytometry was used to test the effects of Allicin on cell cycle. Immunocytochemical technique was used to detect the effects on expression of intracellular apoptotic protein Bax and anti-apoptotic protein Bcl-2 after treatment by Allicin. Results Compared with the control group， different concentrations of Allicin could inhibit the growth of TPC-1 cells， the differences were statistically significant （P < 0.05）; the inhibition rate of Allicin on cells was related to time and concentration. With the increase of Allicin concentration， the proportion of G2/M phase cells was significantly increased， while that of G0/G1 phase cells was decreased. All pairwise comparison of adjacent concentration groups showed statistically significant differences （P < 0.05） except the control group and 10 μg/mL Allicin group （P > 0.05）. Compared with the control group， the intracellular Bcl-2 expression was decreased and Bax expression was increased in every concentration of Allicin treatment group， the differences were statistically significant （P < 0.05）; the expression levels of Bcl-2 and Bax in Allicin treatment groups with different adjacent concentrations were compared， the differences were statistically significant （P < 0.05）. Conclusion Allicin can inhibit the growth of TPC-1 cells of papillary thyroid cancer， and its mechanism may be related to inhibition of cell proliferation by Allicin and the induction of apoptosis by down-regulating Bcl-2 and up-regulating Bax.

[Key words] Allicin; Papillary thyroid carcinoma; Cell cycle; Bcl-2; Bax

甲狀腺癌是一種常見的惡性腫瘤，在我國發病率逐年上升，尤其是在我國沿海地區發病率增長速度更快，女性因受到雌激素的影響，發病率也明顯高于男性[1-2]。甲狀腺癌類型包括未分化癌、乳頭狀癌、濾泡狀癌、髓樣癌。其中乳頭狀癌最為常見，惡性度較低，預后較好。當前醫學界對于甲狀腺癌的主要治療方式為手術切除，患者術后需要服用相關藥物來完成輔助治療，但臨床治療結果表明部分甲狀腺乳頭狀癌在病情發生、發展過程中出現失分化，生長速率加快，進而出現癌細胞轉移等特征，從而影響患者的愈后。因此，尋找新的安全、有效的輔助療法，對提升甲狀腺乳頭狀癌的臨床治療效果，同時保證患者的生存質量，減少患者痛苦具有重要意義。……