Aim: To establish a new method for labeling of procollagen gene probe and the detection of procollagen mRNA expression in HSC.
Methods: According to gene sequences from NCBI Gene Bank, the primers for the amplification of type I, III and IV procollagen genes were designed by OLIGO software, the procollagen genes were amplified by RT-PCR and were labeled by PCR and DIG-dUTP, and the probes were applied to detect procollagen mRNA expression in cultured hepatic stellate cells by in situ hybridization.
Results: The procollagen genes were successfully amplified by the Primers and RT-PCR and were labeled by DIG-dUTP and PCR, the procollagen mRNA expression in cultured hepatic stellate cells was detected by the probes.
Conclusions: A new simplified method for labeling of procollagen genes is successfully found and it can be used for other gene amplification and labeling.