CAJ | 학술논문

目的：观察结肠癌组织及结肠癌细胞中miR-92a表达变化，并探讨其临床意义。方法①选择Ⅱ期合并高危因素及Ⅲ期结肠癌患者75例为研究对象，采用实时荧光定量PCR方法和2－ΔΔCt法测算结肠癌组织miR-92a的相对表达量，分析miR-92 a相对表达量与患者临床病理参数的关系。②取常规培养的结肠癌细胞株HT-29接种于96孔板，设实验组、阴性对照组、脂质体组和对照组，各3个复孔。实验组转染miR-92a抑制剂，阴性对照组转染对照寡核苷酸，脂质体组转染脂质体，对照组不作任何处理，转染后检测miR-92a表达以验证转染效果，转染次日细胞贴壁生长后加50μg／mL奥沙利铂作用24 h，分别在24、48、72 h加入10μL CCK-8溶液，37℃孵育2 h，在酶标仪上读取各组波长为450nm处的吸光度（A）值。转染48 h后加50μg／mL奥沙利铂作用24 h，采用TUNEL法测算各组细胞凋亡率。结果结肠癌组织中miR-92a相对表达量显著高于癌旁正常组织。结肠癌组织中miR-92a的表达水平与患者性别、年龄、肿瘤部位、浸润深度、有无淋巴结转移、脉管癌栓和病理分化程度无关（P均＞0．05），与术后局部复发及发生远处转移有关（ P＜0．05）。 HT-29细胞转染后24、48、72 h实验组A值均低于其余三组（ P均＜0．05），细胞凋亡率高于其余三组（P均＜0．05）。结论结肠癌组织中miR-92a表达水平升高。结肠癌患者术后局部复发及远处转移与结肠癌组织中miR-92a相对表达量有关。抑制结肠癌细胞中miR-92a表达可增加结肠癌细胞化疗敏感性。
목적：관찰결장암조직급결장암세포중miR-92a표체변화，병탐토기림상의의。방법①선택Ⅱ기합병고위인소급Ⅲ기결장암환자75례위연구대상，채용실시형광정량PCR방법화2－ΔΔCt법측산결장암조직miR-92a적상대표체량，분석miR-92 a상대표체량여환자림상병리삼수적관계。②취상규배양적결장암세포주HT-29접충우96공판，설실험조、음성대조조、지질체조화대조조，각3개복공。실험조전염miR-92a억제제，음성대조조전염대조과핵감산，지질체조전염지질체，대조조불작임하처리，전염후검측miR-92a표체이험증전염효과，전염차일세포첩벽생장후가50μg／mL오사리박작용24 h，분별재24、48、72 h가입10μL CCK-8용액，37℃부육2 h，재매표의상독취각조파장위450nm처적흡광도（A）치。전염48 h후가50μg／mL오사리박작용24 h，채용TUNEL법측산각조세포조망솔。결과결장암조직중miR-92a상대표체량현저고우암방정상조직。결장암조직중miR-92a적표체수평여환자성별、년령、종류부위、침윤심도、유무림파결전이、맥관암전화병리분화정도무관（P균＞0．05），여술후국부복발급발생원처전이유관（ P＜0．05）。 HT-29세포전염후24、48、72 h실험조A치균저우기여삼조（ P균＜0．05），세포조망솔고우기여삼조（P균＜0．05）。결론결장암조직중miR-92a표체수평승고。결장암환자술후국부복발급원처전이여결장암조직중miR-92a상대표체량유관。억제결장암세포중miR-92a표체가증가결장암세포화료민감성。
Objective To observe the expression changes in levels of miR-92a in the colon cancer tissues and cells andtoexploreitsclinicalsignificance.Methods ①TherelativeexpressionlevelsofmiR-92ainthetissuesof75pa-tients with stagesⅡwith high risk factors or stagesⅢcolon cancer was detected by real-time q-PCR and was calculated by using the equation 2 -ΔΔCt , and its association with clinicopathological parameter was evaluated.②HT-29 cells were plated onto 96-well plates and were divided into the experimental group, negative control group, liposome group and control group, each group set up three holes.The experimental group was transfected with miR-92a inhibitor, the negative control group was transfected with miR-92a negative control oligonucleotide, and liposome group was transfected liposome.We de-tected the expression changes of miR-92a to evaluated the effects of transfection.Then the cells were treated with 50μg/mL oxaliplatin for 24 h, and at different time points (24, 48 and72 h), the culture medium was removed and replaced with culture medium containing 10μL CCK-8 solution.After incubation at 37℃for 2 h, the absorbance at 450 nm on each well was measured by an enzyme immunoassay analyzer.After the transfection for 48 h and being treated by oxaliplatin for 24 h, the apoptosis rate was detected and measured by TUNEL.Results The expression of miR-92a was significantly higher than that of the normal adjacent mucosa.The expression of miR-92a was not related with gender, age, tumor location, depth of invasion, lymph node metastasis, cancer embolus or tissue differentiation ( all P>0.05) , but was related with lo-cal recurrence and distant metastases (P<0.05).The experimental groups'A value of HT-29 were lower and the apoptosis rate was higher as compared with that of the other three group at different time points (24,48 and 72 h) after transfection (all P<0.05).Conclusions The expression of miR-92a in the colon cancer tissues is increased.The relative expression of miR-92a in the colon cancer tissues is related with local recurrence and distant metastases.Suppressing the expression of miR-92a can increase the chemosensitivity of colon cancer cells.