CAJ | 학술논문

目的:观察MCF-7细胞低表达半胱氨酰白三烯Ⅱ型受体(CysLT2R)对半胱氨酰白三烯(LTC4)介导的细胞生长、侵袭能力、细胞周期和凋亡等生物学行为的影响.方法:针对CysLT2R的已知cDNA序列,设计并体外转录合成4条特异性shRNA,用脂质体介导转染入MCF-7细胞48 h,同时设阴性对照组(1条非特异性序列转染)、正常MCF-7对照组(未转染).Western blot检测干扰后CysLT2R蛋白表达水平下调的变化.应用噻唑蓝比色(MTT)法检测50 nM LTC4作用下细胞株生长能力;应用流式细胞仪检测50 nM LTC4作用下细胞周期和凋亡变化;应用趋化小室检测50 nM LTC4作用下MCF-7细胞侵袭运动活性.结果:与正常MCF-7细胞和阴性对照组比较,MCF-7细胞CysLT2R低表达组的细胞生长增殖无明显变化(P>0.05),干扰后,MCF-7细胞表现出侵袭活性明显增强(P<0.05),细胞周期和凋亡无明显变化.结论:CysLT2R在人乳腺癌MCF-7细胞的侵袭过程中起着一定的作用,CysLT2R有可能是一种新的乳腺肿瘤抑制基因.
목적:관찰MCF-7세포저표체반광안선백삼희Ⅱ형수체(CysLT2R)대반광안선백삼희(LTC4)개도적세포생장、침습능력、세포주기화조망등생물학행위적영향.방법:침대CysLT2R적이지cDNA서렬,설계병체외전록합성4조특이성shRNA,용지질체개도전염입MCF-7세포48 h,동시설음성대조조(1조비특이성서렬전염)、정상MCF-7대조조(미전염).Western blot검측간우후CysLT2R단백표체수평하조적변화.응용새서람비색(MTT)법검측50 nM LTC4작용하세포주생장능력;응용류식세포의검측50 nM LTC4작용하세포주기화조망변화;응용추화소실검측50 nM LTC4작용하MCF-7세포침습운동활성.결과:여정상MCF-7세포화음성대조조비교,MCF-7세포CysLT2R저표체조적세포생장증식무명현변화(P>0.05),간우후,MCF-7세포표현출침습활성명현증강(P<0.05),세포주기화조망무명현변화.결론:CysLT2R재인유선암MCF-7세포적침습과정중기착일정적작용,CysLT2R유가능시일충신적유선종류억제기인.
Objective:The objective of this research is to study the effects of cysteinyl leukotriene receptorⅡ(CysLT2R) on the biological behavior of the MCF-7 human breast carcinoma cell line facilitated by proinflammatory leukotriene C4 (LTC4). Methods:Four small interfering RNAs (shRNA) targeting CysLT2R were transfected into the MCF-7 human breast carcinoma cell line. Western blot assay was used to determine the down-regulation of the protein. The cell cycle changes and apoptosis in the presence of LTC4 were detected using flow cytometry. The proliferation of cells under the impact of 50 nM LTC4 was analyzed using methyl thiazolyl tetrazolium assay. Cell invasion assays were performed on breast cancer cells in the presence of LTC4. Results:No significant change in the cell cycle and apoptosis among the MCF-7 cells (transfected with CysLT2R-shRNA) were observed. The low-expression of CysLT2R had no significant effect on the proliferation of MCF-7 cells. The LTC4-induced cell invasion was evidently increased by the silencing of CysLT2R (P<0.05). Conclusion:Our results show the potential role of CysLT2R in the process of breast cancer cell invasion. CysLT2R may be a new breast carcino ma-suppressor gene.