中国癌症杂志
中國癌癥雜誌
중국암증잡지
CHINA ONCOLOGY
2014年
8期
561-567
,共7页
共刺激分子%B7-H3%食管癌%Eca-109
共刺激分子%B7-H3%食管癌%Eca-109
공자격분자%B7-H3%식관암%Eca-109
Costimulatory molecule%B7-H3%Esophageal cancer%Eca-109
背景与目的：食管肿瘤严重威胁着人类健康，认识食管癌的发生、发展机制和寻找治疗方法已经成为遏制肿瘤的研究热点。近年来，作为B7免疫球蛋白超家族的新成员，共刺激分子B7-H3在多种肿瘤中异常高表达，被认为可能是一种新的肿瘤标志物和潜在的治疗靶点。本研究旨在检测食管癌细胞株TE-1、TE-13、Eca-109细胞中B7-H3的表达，并通过靶向干扰B7-H3基因表达来研究B7-H3对食管癌细胞增殖、侵袭等生物学行为的影响。方法：运用逆转录聚合酶链反应(reverse transcription polymerase chain reaction，RT-PCR)方法检测B7-H3分子在食管癌细胞TE-1、TE-13、Eca-109中的表达。通过LipofectamineTM2000体外转染B7-H3 siRNA、control siRNA至食管癌Eca-109细胞，采用RT-PCR和蛋白质印迹法(Western blot)检测Eca-109细胞中B7-H3 mRNA和蛋白的表达。采用MTT实验、细胞划痕实验、Transwell侵袭小室实验检测B7-H3 siRNA对Eca-109细胞增殖能力、平面迁移能力及体外侵袭能力的影响。结果：共刺激分子B7-H3在食管癌细胞TE-1(0.382±0.008)、TE-13(0.399±0.008)、Eca-109(0.428±0.012)中的表达差异无统计学意义(P>0.05)。转染B7-H3 siRNA后Eca-109细胞B7-H3 mRNA和蛋白表达水平显著低于未转染组(0.1285±0.0002vs 0.5403±0.0013，0.4214±0.0048vs 0.4921±0.0148，P均<0.05)以及空载体转染组(0.1285±0.0002vs 0.5324±0.0007，0.4214±0.0048vs 0.5006±0.0129，P均<0.05)。与对照组细胞相比，转染B7-H3 siRNA后Eca-109细胞的平面迁移能力和侵袭力明显下降(P<0.05)，然而其增殖能力差异无统计学意义(P>0.05)。结论：食管癌细胞TE-1、TE-13、Eca-109均组成性表达B7-H3分子。沉默B7-H3基因表达能明显抑制Eca-109细胞的体外迁移、侵袭能力，提示B7-H3基因可能参与调节食管癌的侵袭转移能力，为免疫治疗提供潜在的治疗靶点。
揹景與目的：食管腫瘤嚴重威脅著人類健康，認識食管癌的髮生、髮展機製和尋找治療方法已經成為遏製腫瘤的研究熱點。近年來，作為B7免疫毬蛋白超傢族的新成員，共刺激分子B7-H3在多種腫瘤中異常高錶達，被認為可能是一種新的腫瘤標誌物和潛在的治療靶點。本研究旨在檢測食管癌細胞株TE-1、TE-13、Eca-109細胞中B7-H3的錶達，併通過靶嚮榦擾B7-H3基因錶達來研究B7-H3對食管癌細胞增殖、侵襲等生物學行為的影響。方法：運用逆轉錄聚閤酶鏈反應(reverse transcription polymerase chain reaction，RT-PCR)方法檢測B7-H3分子在食管癌細胞TE-1、TE-13、Eca-109中的錶達。通過LipofectamineTM2000體外轉染B7-H3 siRNA、control siRNA至食管癌Eca-109細胞，採用RT-PCR和蛋白質印跡法(Western blot)檢測Eca-109細胞中B7-H3 mRNA和蛋白的錶達。採用MTT實驗、細胞劃痕實驗、Transwell侵襲小室實驗檢測B7-H3 siRNA對Eca-109細胞增殖能力、平麵遷移能力及體外侵襲能力的影響。結果：共刺激分子B7-H3在食管癌細胞TE-1(0.382±0.008)、TE-13(0.399±0.008)、Eca-109(0.428±0.012)中的錶達差異無統計學意義(P>0.05)。轉染B7-H3 siRNA後Eca-109細胞B7-H3 mRNA和蛋白錶達水平顯著低于未轉染組(0.1285±0.0002vs 0.5403±0.0013，0.4214±0.0048vs 0.4921±0.0148，P均<0.05)以及空載體轉染組(0.1285±0.0002vs 0.5324±0.0007，0.4214±0.0048vs 0.5006±0.0129，P均<0.05)。與對照組細胞相比，轉染B7-H3 siRNA後Eca-109細胞的平麵遷移能力和侵襲力明顯下降(P<0.05)，然而其增殖能力差異無統計學意義(P>0.05)。結論：食管癌細胞TE-1、TE-13、Eca-109均組成性錶達B7-H3分子。沉默B7-H3基因錶達能明顯抑製Eca-109細胞的體外遷移、侵襲能力，提示B7-H3基因可能參與調節食管癌的侵襲轉移能力，為免疫治療提供潛在的治療靶點。
배경여목적：식관종류엄중위협착인류건강，인식식관암적발생、발전궤제화심조치료방법이경성위알제종류적연구열점。근년래，작위B7면역구단백초가족적신성원，공자격분자B7-H3재다충종류중이상고표체，피인위가능시일충신적종류표지물화잠재적치료파점。본연구지재검측식관암세포주TE-1、TE-13、Eca-109세포중B7-H3적표체，병통과파향간우B7-H3기인표체래연구B7-H3대식관암세포증식、침습등생물학행위적영향。방법：운용역전록취합매련반응(reverse transcription polymerase chain reaction，RT-PCR)방법검측B7-H3분자재식관암세포TE-1、TE-13、Eca-109중적표체。통과LipofectamineTM2000체외전염B7-H3 siRNA、control siRNA지식관암Eca-109세포，채용RT-PCR화단백질인적법(Western blot)검측Eca-109세포중B7-H3 mRNA화단백적표체。채용MTT실험、세포화흔실험、Transwell침습소실실험검측B7-H3 siRNA대Eca-109세포증식능력、평면천이능력급체외침습능력적영향。결과：공자격분자B7-H3재식관암세포TE-1(0.382±0.008)、TE-13(0.399±0.008)、Eca-109(0.428±0.012)중적표체차이무통계학의의(P>0.05)。전염B7-H3 siRNA후Eca-109세포B7-H3 mRNA화단백표체수평현저저우미전염조(0.1285±0.0002vs 0.5403±0.0013，0.4214±0.0048vs 0.4921±0.0148，P균<0.05)이급공재체전염조(0.1285±0.0002vs 0.5324±0.0007，0.4214±0.0048vs 0.5006±0.0129，P균<0.05)。여대조조세포상비，전염B7-H3 siRNA후Eca-109세포적평면천이능력화침습력명현하강(P<0.05)，연이기증식능력차이무통계학의의(P>0.05)。결론：식관암세포TE-1、TE-13、Eca-109균조성성표체B7-H3분자。침묵B7-H3기인표체능명현억제Eca-109세포적체외천이、침습능력，제시B7-H3기인가능삼여조절식관암적침습전이능력，위면역치료제공잠재적치료파점。
Background and purpose:Esophageal cancer is a serious disease threatening human health, and it is very difficult to understand the development mechanism and find the therapeutic methods for esophageal cancer. In recent years, B7-H3, as a new member of B7 immunoregulatory superfamily, overexpressed in multiple tumor types, is considered to be a new tumor marker and potential therapeutic target. This study aimed to detect the expression of B7-H3 in esophageal cancer cell lines TE-1, TE-13, Eca-109 and exploring the effect of B7-H3 siRNA on cell proliferation, migration and invasionin vitro in human esophageal cancer Eca-109 cell line. Methods:The expression of B7-H3 in esophageal cancer cell lines TE-1, TE-13 and Eca-109 were detected by reverse transcription polymerase chain reaction (RT-PCR). B7-H3 siRNA and control siRNA were transfectedin vitro into human esophageal cancer Eca-109 cells using LipofectamineTM 2000. The expressions of B7-H3 mRNA and protein in Eca-109 cells were analyzed by RT-PCR and Western blot. The proliferation, migration and invasion abilities of Eca-109 cells were measured by MTT assay, wound scrape assay and transwell invasion assayin vitro,respectively.Results:All tested cultured esophageal cancer cell lines constitutively expressed B7-H3 mRNA under normal conditions (TE-1 0.382±0.008, TE-13 0.399±0.008, Eca-109 0.428±0.012). After transfection, the expression of B7-H3 mRNA levels decreased in B7-H3 siRNA transfected group, compared with control siRNA transfected group (0.128 5±0.000 2vs 0.532 4±0.000 7,P<0.01) and untransfected group (0.128 5±0.000 2vs 0.540 3±0.001 3,P<0.01), while its protein expression levels were also signiifcantly lower than the control transfection group (0.421 4±0.004 8vs 0.500 6±0.012 9,P<0.05) and untransfected group (0.421 4±0.004 8vs 0.492 1±0.014 8, P<0.05). Compared with control transfected and untransfected cells, Eca-109 cell migration and invasion abilities decreased significantly (P<0.05) by siRNA interference, but no significant difference was observed between their proliferative capacity (P>0.05).Conclusion:All tested esophageal cancer cell lines constitutively express B7-H3 mRNA. B7-H3 siRNA interference inhibits Eca-109 cell migration and invasion abilities. B7-H3 may have a critical role in regulating Eca-109 cell progression.
