本文選題：miR-　切入點(diǎn)：肝癌　出處：《中華腫瘤防治雜志》2017年16期 　論文類型：期刊論文

【摘要】：目的 miR-122是一種肝臟特異性miRNA,miR-122過表達(dá)可以降低肝臟腫瘤細(xì)胞的相關(guān)特性,并增強(qiáng)化療藥物的敏感性,但是在放療方面研究較少。本研究探討miR-122對肝癌細(xì)胞放療敏感性的影響及其可能的機(jī)制。方法通過轉(zhuǎn)染miR-122類似物構(gòu)建HepG2-miR-122mimic細(xì)胞,利用Q-PCR檢測miR-122的表達(dá),通過成克隆分析觀察miR-122對肝癌細(xì)胞放療敏感性的影響,ELISA方法檢測X射線照射后不同時間點(diǎn)細(xì)胞內(nèi)乏氧因子HIF1-α及ROS的表達(dá);蛋白質(zhì)印跡法檢測X射線照射后48h各細(xì)胞凋亡相關(guān)蛋白Bcl-2、Bax及Bim的表達(dá)變化。結(jié)果 Q-PCR檢測顯示,HepG2-miR-122mimic細(xì)胞中miR-122的表達(dá)較HepG2細(xì)胞增加了約8.2倍。成克隆分析顯示,HepG2細(xì)胞轉(zhuǎn)染miR-122mimic后放療敏感性增高,與HepG2組相比,準(zhǔn)域劑量Dq的放療增敏比(SER)為1.52。ELISA分析顯示,HepG2細(xì)胞在0hHIF-1α表達(dá)量為5.20ng/mL。X射線照射后HepG2細(xì)胞中HIF-1α升高,至48h升至最高,為6.69ng/mL。而HepG2-miR-122mimic細(xì)胞在0h的表達(dá)量為4.7ng/mL,X射線照射后HIF-1α的含量進(jìn)一步下降,至48h下降至3.41ng/mL。48h時miR-122 mimic轉(zhuǎn)染情況對HIF-1α的表達(dá)差異有統(tǒng)計學(xué)意義,F=70.819,P0.001;X射線暴露對HIF-1α的表達(dá)差異有統(tǒng)計學(xué)意義,F=132.436,P0.001;miR-122mimic的轉(zhuǎn)染情況及X射線暴露之間存在交互作用,F=4.290,P=0.039。ROS含量在X射線照射后表現(xiàn)為輕度升高,在0h HepG2組和HepG2-miR-122mimic組ROS表達(dá)量分別為116.21和121IU/mL,X射線照射后ROS進(jìn)一步升高,至48h升至最高,分別為130和192IU/mL。48h時miR-122mimic轉(zhuǎn)染情況對ROS的表達(dá)差異有統(tǒng)計學(xué)意義,F=98.189,P0.001;X射線暴露對ROS的表達(dá)差異有統(tǒng)計學(xué)意義,F=145.373,P0.001;miR-122mimic的轉(zhuǎn)染情況及X射線暴露之間存在交互作用,F=6.548,P=0.012。蛋白質(zhì)印跡法檢測結(jié)果顯示,HepG2-miR-122mimic細(xì)胞中BCL-2的表達(dá)量為HepG2的56.32%,X射線照射后48hHepG2細(xì)胞中BCL-2的表達(dá)量降至原來的55.01%,而HepG2-miR-122mimic細(xì)胞則降至23%。HepG2-miR-122mimic細(xì)胞中Bax和Bim的表達(dá)量分別為HepG2細(xì)胞的170.21%和140.35%,X射線照射后,HepG2細(xì)胞中Bax和Bim的表達(dá)量分別升至HepG2的220.12%和178.34%,而HepG2-miR-122mimic細(xì)胞中,Bax和Bim的表達(dá)量則分別升至HepG2細(xì)胞的240%和260%。miR-122mimic轉(zhuǎn)染情況對BCL-2、Bax和Bim的表達(dá)差異均有統(tǒng)計學(xué)意義,均P0.01。X射線暴露對BCL-2、Bax及Bim的表達(dá)差異均有統(tǒng)計學(xué)意義,均P0.01。miR-122mimic轉(zhuǎn)染情況及X射線暴露對Bcl-2、Bim及Bax的表達(dá)均不存在交互作用。結(jié)論 miR-122可以上調(diào)肝癌細(xì)胞HepG2的放射敏感性,其機(jī)制可能與調(diào)節(jié)HIF-1α和ROS及凋亡相關(guān)蛋白有關(guān)。
[Abstract]:Objective miR-122 is a liver-specific miRNA-miR-122 overexpression that can reduce the related characteristics of liver tumor cells and enhance the sensitivity of chemotherapeutic drugs. In this study, we studied the effect of miR-122 on the radiosensitivity of hepatoma cells and its possible mechanism. Methods HepG2-miR-122mimic cells were constructed by transfection of miR-122 analogues, and the expression of miR-122 was detected by Q-PCR. The effects of miR-122 on the radiosensitivity of hepatoma cells were observed by clone analysis. The expression of hypoxia factor HIF1- 偽 and ROS were detected by Elisa at different time points after X-ray irradiation. The expression of apoptosis-related protein Bcl-2 Bax and Bim in HepG2-miR-122 mimic cells was detected by Western blotting at 48 h after irradiation. Results the expression of miR-122 in HepG2-miR-122 mimic cells was increased by about 8.2-fold compared with that in HepG2 cells. Clone analysis showed that miR-122 expression in HepG2 cells was about 8.2-fold higher than that in HepG2 cells. After transfection of miR-122mimic, the sensitivity of radiotherapy was increased. Compared with HepG2 group, 1.52.ELISA analysis showed that the expression of 0hHIF-1 偽 in HepG2 cells increased after 5.20ng/mL.X irradiation, and reached the highest level at 48 h. The expression of HIF-1 偽 in HepG2-miR-122mimic cells was 4.7ng / mLX irradiation at 0 h, and the content of HIF-1 偽 decreased further after X-ray irradiation. There was significant difference in the expression of HIF-1 偽 between miR-122 mimic transfection and 3.41ng/mL.48h at 48h. There was a significant difference in the expression of HIF-1 偽 after X-rays exposure to miR-122 mimic. There was a significant difference in the expression of HIF-1 偽. There was a significant difference in the transfection of HIF-1 偽 and the interaction between F4.290P0.039.Ros content in X-ray exposure. A slight increase was observed after irradiation. The expression of ROS in 0 h HepG2 group and HepG2-miR-122mimic group were 116.21 and 121 IU / mL X ray irradiation respectively, and ROS increased further, and reached the highest level at 48 h. There were significant differences in the expression of ROS between miR-122mimic transfection and 192IU/mL.48h respectively. There was a significant difference in the expression of ROS by X-Ray exposure to FN 98.189 and P0.001. There was a significant difference in the transfection of FN145.373 and P0.001miR-122 mimic and the interaction between ROS and X-ray exposure. Western blotting was used to detect the expression of ROS. The results showed that the expression of BCL-2 in HepG2-miR-122 mimic cells was 56.32% of that of HepG2. After X-ray irradiation, the expression of BCL-2 in 48hHepG2 cells decreased to 55.01%, while that of Bax and Bim in HepG2-miR-122mimic cells decreased to 170.21% and 140.35% of HepG2 cells, respectively. The expression of Bax and Bim in HepG2 cells increased to 220.12% and 178.34% of HepG2, respectively, while the expression of HepG2-miR-122mimic cells increased to 240% of HepG2 cells and 240% of 260%.miR-122mimic transfection, respectively. There were significant differences in the expression of BCL-2nBax and Bim in HepG2 cells. There was significant difference in the expression of BCL-2nBax and Bim by P0.01. There was no interaction between P0.01.miR-122mimic transfection and X-ray exposure to Bcl-2mBim and Bax. Conclusion miR-122 can up-regulate the radiosensitivity of hepatoma cells to HepG2. The mechanism may be related to the regulation of HIF-1 偽 and ROS and apoptosis-related proteins.
【作者單位】： 江蘇大學(xué)附屬人民醫(yī)院腫瘤放療科;
【基金】：[基金項(xiàng)目]鎮(zhèn)江市科技支撐計劃-社會發(fā)展(SH2014040)
【分類號】：R730.55;R735.7

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