Nodosin通過抑制mTOR通路和激活自噬誘導(dǎo)結(jié)直腸癌細(xì)胞凋亡

背景：結(jié)直腸癌是消化道常見的惡性腫瘤，現(xiàn)有的治療方法和藥物具有毒副反應(yīng)大等缺點(diǎn)，因此迫切需要研發(fā)出高效低毒的新型治療藥物。目的：探討Nodosin誘導(dǎo)結(jié)直腸癌細(xì)胞凋亡的機(jī)制。方法：體外培養(yǎng)結(jié)直腸癌細(xì)胞株SW480、HCT116，以0、5、10和20 μmol/L Nodosin處理細(xì)胞24 h，或以10 μmol/L Nodosin處理細(xì)胞0、12、24和48 h。以CCK?8法檢測細(xì)胞活性，蛋白質(zhì)印跡法檢測PARP1、mTOR、p?mTOR、LC3BⅡ蛋白表達(dá)。SW480、HCT116細(xì)胞以5 mmol/L自噬抑制劑3?甲基腺嘌呤（3?MA）預(yù)處理4 h以后，與10 μmol/L Nodosin培養(yǎng)24 h，蛋白質(zhì)印跡法檢測PARP1、LC3BⅡ蛋白表達(dá)。結(jié)果：Nodosin能呈濃度依賴性和時(shí)間依賴性地抑制SW480、HCT116細(xì)胞活性，凋亡相關(guān)蛋白cleaved PARP1和自噬相關(guān)蛋白LC3BⅡ表達(dá)升高，p?mTOR表達(dá)受抑。自噬抑制劑3?MA預(yù)處理可抑制Nodosin誘導(dǎo)的凋亡相關(guān)蛋白cleaved PARP1和自噬相關(guān)蛋白LC3BⅡ表達(dá)。結(jié)論：Nodosin可通過抑制mTOR通路和激活自噬誘導(dǎo)結(jié)直腸癌細(xì)胞凋亡。

關(guān)鍵詞 Nodosin； 自噬； 結(jié)直腸腫瘤； 細(xì)胞凋亡

Nodosin Induces Apoptosis of Colorectal Cancer Cells by Inhibiting mTOR Pathway and Activating Autophagy ZHANG Hong， CAI Shasha， XIE Chunqin， YU Changfa， FANG Han. Department of Laboratory Medicine， Taizhou First People′s Hospital， Taizhou， Zhejiang Province （318020）

Background： Colorectal cancer is a common malignant tumor in the digestive tract. The existing treatment methods and drugs have disadvantages such as significant toxicity and side effects， so there is an urgent need to develop new drugs with high efficiency and low toxicity. Aims： To investigate the potential mechanism of Nodosin in inducing apoptosis of colorectal cancer cells. Methods： Colorectal cancer cell lines SW480 and HCT116 were cultured with 0， 5， 10 and 20 μmol/L Nodosin for 24 hours， or cultured with 10 μmol/L Nodosin for 0， 12， 24 and 48 hours. The cell viability was measured by CCK?8 assay， and the protein expressions of PARP1， mTOR， p?mTOR and LC3BⅡ were detected by Western blotting. After pretreated with the autophagy inhibitor 3?methyladenine （3?MA） 5 mmol/L for 4 hours， SW480 and HCT116 cells were cultured with 10 μmol/L Nodosin for 24 hours. The protein expressions of PARP1 and LC3BⅡ were detected by Western blotting. Results： Nodosin could decrease the cell viability of SW480 and HCT116 cells in a dose? and time?dependent manner， increase the expressions of apoptosis?related protein cleaved PARP1 and autophagy?related protein LC3BⅡ， and inhibit the protein expression of p?mTOR. Pretreatment of the autophagy inhibitor 3?MA could inhibit the expressions of apoptosis?related protein cleaved PARP1 and autophagy?related protein LC3BⅡ induced by Nodosin. Conclusions： Nodosin can induce apoptosis of colorectal cancer cells by inhibiting mTOR pathway and activating autophagy.

Key words Nodosin; Autophagy; Colorectal Neoplasms; Apoptosis

結(jié)直腸癌（colorectal cancer）是消化道常見的惡性腫瘤，其發(fā)病率逐年增高，已成為全球四大致死性惡性腫瘤之一，每年約有90萬例患者死亡[1]。目前的治療手段包括放療、化療、靶向治療、免疫治療等[2]。雖然化療藥物可以降低腫瘤復(fù)發(fā)率和轉(zhuǎn)移率，但不良反應(yīng)多，且容易產(chǎn)生耐藥導(dǎo)致治療失敗[3]；靶向治療這一新型抗癌手段雖然得到廣泛關(guān)注和認(rèn)可，但仍存在患者依從性差的問題；免疫療法對多種癌癥具有卓越的療效，但在結(jié)直腸癌的治療上因患者個(gè)體差異而有明顯不同[2?3]。因此，對于結(jié)直腸癌的治療，仍需要探尋新的高效低毒的治療藥物。

Nodosin是從藥用植物溪黃草中提取的二萜類化合物，已被證實(shí)具有多種生物學(xué)功能[4]。……