RNA干扰沉默RhoC基因抗炎性乳腺癌侵袭的研究

Experimental study on inhibiting invasion of inflammatory breast cancer cell lines SUM149 and SUM190 by silencing RhoC gene with the help of RNA interference

目的观察利用RNA干扰（RNAi）技术沉默炎性乳腺癌（IBC）细胞SUM149和SUM190 RhoC基因对肿瘤细胞侵袭的影响。方法人炎性乳腺癌细胞按均衡随机方法分为空白对照组、阴性脂质体组、RhoC-小干扰RNA（siRNA）转染组，人炎性乳腺癌细胞SUM149和SUM190中转染RhoC-siRNA，利用显微镜分析转染效率。转染后48 h，应用反转录-聚合酶链反应（RT-PCR）检测RhoC mRNA的表达水平，应用Western blot检测RhoC蛋白的表达水平。利用Transwell小室检测转染后肿瘤细胞的侵袭能力。结果应用RhoC-siRNA转染人炎性乳腺癌细胞SUM149和SUM190，转染效率可以达到80%。在SUM149细胞中，阴性脂质体组与RhoC-siRNA转染组RhoC蛋白的平均表达水平分别为0.897±0.376、0.375±0.164，差异有统计学意义（t＝6.274，P＝0.045）。在SUM190细胞中，阴性脂质体组与RhoC-siRNA转染组RhoC蛋白的平均表达水平分别为0.902±0.386、0.381±0.166，差异有统计学意义（t＝6.891，P＝0.028）。在侵袭实验中，SUM149细胞RhoC-siRNA转染组和阴性脂质体组穿膜细胞数分别为（85.6±12.7）、（340.5±43.4）个，差异有统计学意义（t＝12.275，P＝0.034）。SUM190细胞RhoC-siRNA转染组和阴性脂质体组穿膜细胞数分别为（92.8±15.6）、（345.9±47.7）个，差异有统计学意义（t＝11.769，P＝0.022）。结论应用RhoC-siRNA转染人炎性乳腺癌细胞SUM149和SUM190，可以有效抑制肿瘤细胞的侵袭。

ObjectiveIn this study, we silence RhoC gene in inflammatory breast cancer （IBC） SUM149, SUM190 cell lines and explore its effects on cell invasion of IBC by RNA interference （RNAi） technology so as to provide experimental principal for target gene therapy of IBC.MethodsHuman IBC cells were divided blank group, control siRNA group, RhoC-siRNA group by balance randomization. RhoC-siRNA was used to transfect human IBC SUM149, SUM190 cell lines. Microscopy was applied to analyze transfection efficiency. After transfecting for 48 hours, reverse transcriptase-polymerase chain reaction （RT-PCR） was made to detect mRNA expression levels of RhoC. Western blotting was also used to analyze the expression level of RhoC protein. After transfecting for 48 hours, the ability to invasion was analyzed by transwell hole.ResultsWe used siRNA to transfect human IBC SUM149, SUM190 cell lines and found that transfection efficiency can exceed to 80%. After we transfected for 48 hours, the expression levels of RhoC protein in IBC SUM149 cell lines 0.375±0.164 in RhoC siRNA group were significantly lower than those 0.897±0.376 in control siRNA group （t=6.274, P=0.045）. The expression levels of RhoC protein in IBC SUM190 cell lines 0.381±0.166 in RhoC siRNA group were significantly lower than those 0.902±0.386 in control siRNA group （t=6.891, P=0.028）. However, there were no obvious differences between in blank group and control siRNA group. After we transfected for 48 hours, in IBC SUM149 cell lines, the ability to invasion 85.6±12.7 in RhoC siRNA group was markedly lower than that 340.5±43.4 in control siRNA group （t=12.275, P=0.034）. In IBC SUM190 cell lines, the ability to invasion 92.8±15.6 in RhoC siRNA group was markedly lower than that 345.9±47.7 in control siRNA group （t=11.769, P=0.022）.ConclusionRhoC is known as an important role in the carcinogenesis of IBC. With the help of RNAi technology, we can greatly inhibit the invasion of IBC SUM149, SUM190 cell lines by silencing RhoC gene. RNAi technology perhaps becomes an important gene therapy tool for IBC.