免疫球蛋白在结肠癌HT-29细胞中的表达及其生物学活性探讨

Immunoglobulin expression in colon cancer cell line HT-29 and its biological activities

目的明确免疫球蛋白(Ig)在人大肠癌传代细胞系HT-29细胞中的表达情况,并探讨其对肿瘤细胞生物学活性的影响。方法用RT-PCR方法检测Ig重链可变区转录本在HT-29细胞中的表达,并构建含有HT-29特异的CDR3片段的反义重组载体,电穿孔法将其转染入HT-29细胞,观察其对HT-29细胞生长增殖及凋亡等基本生命活动的影响。结果在HT-29细胞内检测到人Ig重链转录本,且测序显示为单一性克隆;含有HT-29特异的CDR3片段的反义重组载体能够有效地抑制HT-29细胞Ig分子的表达,且能够有效的诱导细胞的凋亡(P<0.01)和生长抑制(P<0.05)。结论肿瘤细胞来源的Ig可促进肿瘤细胞的生存和生长。

Objective To determine the expression of immunoglobulins in HT-29 cells （an established colon cancer cell line, and exphore their effect on the biological activities of the cancer cells. Methods The transcripts of variable regions of immunoglobulin heavy chains in HT-29 cells were detected by RT-PCR. Antisense CDR3 （specific to HT-29 ）-pIRES 1 neo vector was constructed, then transfected into HT-29 cells by electroporation. Progrannned cell death and growth inhibition of HT-29 cells were detected by FCM and MTT,respectively. Results The transcripts of Ig heavy chain （VH CDR3 region） were expressed in HT-29 cells. Moreover, they showed a monoclonal characteristic after being sequenced. After transfection of the antisense vector of CDR3 （ specific to HT-29 ） -pIRES 1 neo, expression level of Ig in HT-29 cells was significantly decreased, and growth inhibition （ P 〈 0.05 ） and apoptosis （ P 〈 0.01 ） were induced. Conclusion These results suggest that tumor derived Ig could promote the survival and growth of tumor cells.