摘要
目的构建受1α,25二羟维生素D3调控的雌激素受体α表达载体(VDRE-Tk-ERα),并观察其对雌激素受体阴性乳腺癌细胞的增殖抑制效应。方法利用PCR法得到含有4个拷贝VDRE和Tk启动子的序列并用其替换掉雌激素受体α载体的CMV启动子从而得到VDRE-Tk-ERα表达载体。利用脂质体法将其转染入雌激素受体阴性乳腺癌细胞中并利用MTT法观察其对乳腺癌细胞的抑制效应。结果一定剂量的1α,25二羟维生素D3能够通过VDRE调控下游报告基因的表达,同时在加入他莫西芬后能够显著抑制转染了VDRE-Tk-ERα表达载体的雌激素受体阴性乳腺癌细胞的增殖活性。结论通过该方法能够显著恢复和增强雌激素受体阴性乳腺癌细胞对他莫西芬和1α,25二羟维生素D3的化疗敏感性。
Objective To construct ERα expression vector containing VDRE-Tk and evaluate its inhibitory effect on ERα-negative breast cancer cells under induction of 1α, 25 dihydroxyvitamin D3. Methods The 4×VDRE-Tk sequence of VDRE-pGL3 reporter vector was cloned into ERα/pcDNA3.1^＋ vector, and then transfected into MDA-MB-231 breast cancer cells. Expression ERα was induced by 1α, 25 dihydroxyvitamin D3. The effect of ERα expression on responsiveness of MDA-MB-231 breast cancer cells to Tamoxifen were determined. Results 1α, 25 dihydroxyvitamin D3 could induce expression of reporter gene downstream of VDRE-Tk in a dose-dependent manner. Tamoxifen and 1α, 25 dihydroxyvitamin D3 could synergistically inhibited the proliferation of MDA-MB-231 breast cancer cells transfected with VDRE-Tk-ERα expression vector in a time- and dose-dependent manner. Conclusion Transfection of VDRE-Tk-ERα expression vector into ERα-negative breast cancer cells could effectively recover its responsiveness to Tamoxifen under the induction of 1α, 25 dihydroxyvitamin D3.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2006年第2期155-157,161,共4页
Immunological Journal
基金
国家自然科学基金资助项目(30371217)
