现代肿瘤医学
現代腫瘤醫學
현대종류의학
JOURNAL OF MODERN ONCOLOGY
2014年
8期
1770-1774
,共5页
傅韵%蒋蓓琦%夏晨%武羿%王奕静%成小林%李正东%庄志刚
傅韻%蔣蓓琦%夏晨%武羿%王奕靜%成小林%李正東%莊誌剛
부운%장배기%하신%무예%왕혁정%성소림%리정동%장지강
乳腺癌%迁移%侵袭%细胞间隙蛋白 43%细胞间隙连接通讯
乳腺癌%遷移%侵襲%細胞間隙蛋白 43%細胞間隙連接通訊
유선암%천이%침습%세포간극단백 43%세포간극련접통신
breast cancer%migration%invasion%Connexin43%GJIC
目的：探讨 Cx43对乳腺癌侵袭、迁移以及细胞间隙通讯功能的影响。方法：用脂质体转染法将 Cx43质粒、Cx43 siRNA 及阴性对照转染入人乳腺癌细胞株 MDA - MB -231。细胞黏附实验、Transwell 试验检测细胞黏附、侵袭和迁移能力。分子生物学方法检测 MMP -2、MMP -9、BRMS -1的表达。荧光染料传输实验检测不同 Cx43表达的细胞间隙通讯的功能。结果：细胞黏附实验结果表明相较于野生型细胞，Cx43 siRNA 组黏附能力（0.43±0.07）降低（t =20.801，P =0.002），Cx43质粒组黏附能力（1.63±0.26）增高（t =5.917，P =0.027）。Transwell 实验表明相较野生组，Cx43质粒组侵出小室的细胞数增加（侵袭：1.25±0.04，t =16.339，P =0.004；迁移：1.33±0.02，t =22.770，P =0.002），而 Cx43 siRNA 组侵出小室的细胞数减少（迁移：0.84±0.04，t =11.139，P =0.008；侵袭：0.79±0.04，t =5.743，P =0.029）。分子生物学检测发现 Cx43质粒组 MMP -2、MMP -9表达增高，BRMS -1表达降低，而 Cx43 siRNA 组 MMP -2、MMP -9表达降低，BRMS -1表达增高。染料传输实验证实相较野生组传输能力，Cx43质粒组传输作用增强；Cx43 siRNA 组细胞间的传输作用减弱。结论：Cx43对乳腺癌细胞侵袭及迁移能力存在正调控。这一调控能力可能通过 GJIC 实现。
目的：探討 Cx43對乳腺癌侵襲、遷移以及細胞間隙通訊功能的影響。方法：用脂質體轉染法將 Cx43質粒、Cx43 siRNA 及陰性對照轉染入人乳腺癌細胞株 MDA - MB -231。細胞黏附實驗、Transwell 試驗檢測細胞黏附、侵襲和遷移能力。分子生物學方法檢測 MMP -2、MMP -9、BRMS -1的錶達。熒光染料傳輸實驗檢測不同 Cx43錶達的細胞間隙通訊的功能。結果：細胞黏附實驗結果錶明相較于野生型細胞，Cx43 siRNA 組黏附能力（0.43±0.07）降低（t =20.801，P =0.002），Cx43質粒組黏附能力（1.63±0.26）增高（t =5.917，P =0.027）。Transwell 實驗錶明相較野生組，Cx43質粒組侵齣小室的細胞數增加（侵襲：1.25±0.04，t =16.339，P =0.004；遷移：1.33±0.02，t =22.770，P =0.002），而 Cx43 siRNA 組侵齣小室的細胞數減少（遷移：0.84±0.04，t =11.139，P =0.008；侵襲：0.79±0.04，t =5.743，P =0.029）。分子生物學檢測髮現 Cx43質粒組 MMP -2、MMP -9錶達增高，BRMS -1錶達降低，而 Cx43 siRNA 組 MMP -2、MMP -9錶達降低，BRMS -1錶達增高。染料傳輸實驗證實相較野生組傳輸能力，Cx43質粒組傳輸作用增彊；Cx43 siRNA 組細胞間的傳輸作用減弱。結論：Cx43對乳腺癌細胞侵襲及遷移能力存在正調控。這一調控能力可能通過 GJIC 實現。
목적：탐토 Cx43대유선암침습、천이이급세포간극통신공능적영향。방법：용지질체전염법장 Cx43질립、Cx43 siRNA 급음성대조전염입인유선암세포주 MDA - MB -231。세포점부실험、Transwell 시험검측세포점부、침습화천이능력。분자생물학방법검측 MMP -2、MMP -9、BRMS -1적표체。형광염료전수실험검측불동 Cx43표체적세포간극통신적공능。결과：세포점부실험결과표명상교우야생형세포，Cx43 siRNA 조점부능력（0.43±0.07）강저（t =20.801，P =0.002），Cx43질립조점부능력（1.63±0.26）증고（t =5.917，P =0.027）。Transwell 실험표명상교야생조，Cx43질립조침출소실적세포수증가（침습：1.25±0.04，t =16.339，P =0.004；천이：1.33±0.02，t =22.770，P =0.002），이 Cx43 siRNA 조침출소실적세포수감소（천이：0.84±0.04，t =11.139，P =0.008；침습：0.79±0.04，t =5.743，P =0.029）。분자생물학검측발현 Cx43질립조 MMP -2、MMP -9표체증고，BRMS -1표체강저，이 Cx43 siRNA 조 MMP -2、MMP -9표체강저，BRMS -1표체증고。염료전수실험증실상교야생조전수능력，Cx43질립조전수작용증강；Cx43 siRNA 조세포간적전수작용감약。결론：Cx43대유선암세포침습급천이능력존재정조공。저일조공능력가능통과 GJIC 실현。
To explore the effects of Cx43 on the invasion,migration and gap junctional intercellular communication(GJIC)of breast cancer. Methods:The Cx43 plasmid,inhibitors and negative controls were transfected into human breast cancer cell line MDA - MB - 231 by liposome. The motility of cells was evaluated by adhesion and Transwell assay. MMP - 2,MMP - 9 and BRMS - 1 were detected by both qPCR and Western blot. A scrape - loading dye transfer assay was used to evaluate the function of GJIC. Results:Compared to the wild type,adhesion ratio of Cx43 siRNA group(0. 43 ± 0. 07)was significantly lower(t = 20. 801,P = 0. 002),and that of Cx43 plasmid group (1. 63 ± 0. 26)was higher(t = 5. 917,P = 0. 027)by adhesion assay. Transwell tests showed the migration and inva-sion of Cx43 plasmid group was significantly increased( invasion:1. 25 ± 0. 04,t = 16. 339,P = 0. 004;migration:1. 33 ± 0. 02,t = 22. 770,P = 0. 002),but that of Cx43 siRNA group was significantly reduced( migration:0. 84 ± 0. 04,t = 11. 139,P = 0. 008;invasion:0. 79 ± 0. 04,t = 5. 743,P = 0. 029). The expression of MMP - 2 and MMP - 9 were enhanced and the expression of BRMS - 1 was reduced in Cx43 plasmid group detected by both qPCR and west-ern blot and vice versa in Cx43 siRNA group. Compared to the wild type,GJIC was enhanced in Cx43 plasmid group and reduced in Cx43 siRNA group by a scrape - loading dye transfer assay. Conclusion:Cx43 might have a positive control on invasion and migration of breast cancer cell by GJIC.
