The Expression Of VEGFRs And Endostatin In Hemangioma And The Application Of DDRT-PCR

Object:To evaluate the gene expression of VEGFRs and endostatin in hemangioma and the possible relationship between them in two groups (hemangioma and great saphenous vein), the mRNA level in the specimens was detected by Semi-quantitative RT-PCR (Sq RT-PCR) analysis. DDRT-PCR was used to detect the possible differential expressed gene between hemangioma and skin from the same patient. Method:The mRNA level of Flt-1, KDR and endostatin was detected in 15 cases of hemangioma (case group) and lOcases of great saphenous vein (control group). Total RNA was abstracted from the tissue by UNIQ-10 total RNA purification kit, and then analyzed by semi-quantitative reverse transcription and polymerase chain reaction (Sq RT-PCR) method. The brightness of the target band produced by Sq RT-PCR was detected by image analysis system .The signal ratio of the tareget band to β -actin was then analyzed. DDRT-PCR was used to detected the possible differential expressed gene between hemangioma and skin from the same patient.The possible differential expressed gene was then isolated , purified, re-amplified and sequenced. Result:The result showed that the mRNA level of the Flt-1 was 0.6776 ± 0.0286, significantly (p<0.05) higher than those in the control group(0.4668±0.0312); level of the KDR is 0.6303 ±0.0249 , significantly (p<0.05) higher than those in the control group(0.4284 ±0.026) too.There were no statistically significant difference between KDR and Flt-1 in the case group (p>0.05) and the control group(p>0.05). Endostatin was detected in so few cases of two groups that it was difficult to be evaluated. Several possible differential expressed bands were isolated by DDRT-PCR and one of them was sequenced. Conclusion:It is obviously that the expressions of the KDR and Flt-1 were both up-regulated in hemangioma. This finding indicates that VEGF may play a very important role in the progress of hemangioma . And endostatin may be not as important as we expectedin the progress of hemangioma. In the end, DDRT-PCR method proved to be successful and efficient in the detection of differential expressed gene in hemangioma, although with some disadvantages.