Expression And Localization Of SPINK6 In Different Cell Lines

Aims:Human serine protease inhibitor kazal-type 6(SPINK6) is one of several genes which had a huge different expression between the liver cancer cell line and its Parvovirus H-1 resistant cell line while we used microarray to analyze the genes’ expression profile of these two cell lines. But its expression in other cells was not elucidated. The aim of this study was to analyze the expression of SPINK6 in cancer cell lines and normal cell lines, and to research its sub-cellular localizations which initially reveal its mechanism.Methods:Total RNA was extracted from several cell lines using TRIZOL. After reverse transcriptase, we researched the expression of SPINK6 through realtime-PCR in both tumor cell lines and normal cell lines. Furthermore, we transfected several cell lines with the constructed pEGFP-Nl-SPINK6 vector and observe the sub-cellular locations of SPINK6. DAPI was used to tag the nucleus. ER-Tracker Red was used to tag the endoplasmic reticulum. Lyso-Tracker Red was used to tag the lysosome. Dil was used to tag the cell membrane.Results:By realtime-PCR, we found that the expression of SPINK6 in cancer cell lines was lower than that in normal cell lines. The result was said that SPINK6 may be a potential tumor suppressor. We also found that SPINK6 was position in the endoplasmic reticulum and the plasma membrance which may reveal that it play a role throughout the cell membrane.