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目的利用超声靶向转染缺氧诱导因子干扰基因(HIF-1αshRNA),观察其对肝癌的治疗效果。方法构建Wistar大鼠肝癌种植瘤模型。随机将大鼠分为两组:假手术组,开腹后不进行超声辐照治疗;超声靶向基因转染(UTMD)组,开腹后将基因与微泡混合后经尾静脉匀速注入,同时进行超声靶向辐照。分别于治疗后第3天、第7天、第14天行超声造影检测肿瘤大小的变化(计算公式为体积=长×宽×高×π/6),qPCR检测HIF-1αmRNA的变化,Western和免疫组化检测肿瘤细胞内HIF-1α蛋白的表达。结果治疗前两组的肿瘤大小差异无统计学意义。治疗后第3天两组肿瘤均有一定的增大,两组肿瘤大小差异有统计学意义(P<0.05);治疗后第7天,假手术组肿瘤继续增大,UTMD组肿瘤大小保持稳定,两组间差异有统计学意义(P<0.05);治疗后第14天,假手术组肿瘤仍继续增大,UTMD组肿瘤开始缩小,两组间差异有统计学意义(P<0.05)。与假手术组相比,各时间点UTMD组HIF-1αmRNA表达均明显降低,HIF-1α蛋白水平明显下调。结论 UTMD介导HIF-1αshRNA基因转染能有效干扰HIF-1α的表达,使肿瘤组织缺氧、缺血坏死并诱发凋亡,达到抑制肿瘤生长、甚至治愈肿瘤的目的。
OBJECTIVE: To transduce hypoxia inducible factor interfering gene (HIF-1α shRNA) by ultrasound and observe its therapeutic effect on hepatocellular carcinoma. Methods Wistar rat hepatoma tumor model was constructed. The rats were randomly divided into two groups: sham operation group, no ultrasonic radiation treatment after laparotomy; ultrasound-targeted gene transfection (UTMD) group, the gene was mixed with microvesicles after laparotomy, At the same time ultrasound irradiation. The changes of tumor size (calculated as volume = length × width × height × π / 6) were detected by contrast-enhanced ultrasound on the third, seventh and fourteenth day after treatment. The changes of HIF-1αmRNA were detected by qPCR. Western and Immunohistochemical detection of HIF-1α protein expression in tumor cells. Results There was no significant difference in tumor size between the two groups before treatment. On the third day after treatment, the tumors in both groups increased to a certain extent (P <0.05). On the seventh day after treatment, the tumors in the sham-operated group continued to increase and the size of the tumors in the UTMD group remained stable (P <0.05). On the 14th day after treatment, the tumors in the sham operation group continued to increase, and the tumors in the UTMD group began to decrease. The difference between the two groups was statistically significant (P <0.05). Compared with the sham operation group, the expression of HIF-1αmRNA in UTMD group was significantly decreased at each time point, and the protein level of HIF-1α was significantly down-regulated. Conclusion UTMD-mediated HIF-1αshRNA gene transfection can effectively interfere with the expression of HIF-1α, make the tumor tissue hypoxia, necrosis and induce apoptosis, and achieve the goal of inhibiting tumor growth and even curing the tumor.