[The effect of monocarboxylate transporter gene on the regulation of pHi and growth character in cancer cells].

OBJECTIVE: To study the influence of the first subtype of monocarboxylate transporter (MCT1) gene on pHi regulation, lactate transport and cell growth in tumor cells.
METHODS: (1) With RT-PCR technique, MCT1 cDNA fragment was cloned from human lung cancer cells A549, and the cloned fragment MCT1 was reversely inserted into the vector pLXSN to acquire antisense expression recombinant vector pLXSN-MCT1. (2) pLXSN, pLXSN-MCT1 were respectively introduced into the A549 cells by electroporation. The transfected A549 cell resistant to G418 drug was selected as positive clones and proved by PCR. The changes of intracellular pH and lactate in the transfected A549 cells were detected by spectrophotometric method. Cell growth was studied by cell growth curve.
RESULTS: (1) The cloned fragment was in the length of 640 bp and successfully bound to pLXSN. It was also proved to be the objective one by DNA sequencing. (2) Intracellular pH and lactate were remarkably decreased in the cells transfected pLXSN-MCT1, comparing to A549 cells without transfection (P < 0.001). The growth of A549 cells transfected pLXSN-MCT1 was also inhibited remarkably.
CONCLUSION: MCT1 gene could play an important role in pHi regulation, lactate transport and cell growth in tumor cells.