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- _3 value "Overexpression of PrPC {PRNP} enhances tumor cell proliferation, cellular transformation, and tumorigenesis Our previous work has shown that PrPC could be detected in several human gastric cancer cell lines. Relatively lower PrPC expression was found in SGC7901 cells, but higher expression of PrPC was noted in AGS (12) . To show the differential effects of PrPC expression on cell proliferation, PRNP gene was stably transfected into human gastric cancer cell SGC7901 or was blocked by RNAi in AGS cells, respectively. After cell transfection and antibiotic selection for more than 2 mo, multiple drug-resistant clones were selected and the expression of PrPC in the cells was confirmed by Western blot (Fig. 1 A).MTT assay, plate, and soft agar colony formation assays were done to examine the effect of PrPC on cell proliferation and cell transformation in vitro. Compared with empty vector-transfected cells and parental cells, PrPC-transfected cells showed significantly increased rate of cell proliferation as shown by MTT assay (Fig. 1B ). Anchorage-independent growth is one of the important characteristics of in vitro tumor growth; therefore, we examined whether up-regulation of PrPC expression could promote SGC7901 cell growth in soft agar and plate. As shown in Fig. 1C , PrPC transfection resulted in a marked accumulation of growth of SGC7901 in colony formation assay, with average increase rates of 65.1% and 58.6% in soft agar and plate assay, respectively. Down-regulation of PrPC by RNAi inhibited the colony formation both in soft agar and in plate. To confirm this effect in vivo, we injected subcutaneously PrPC-expressing SGC7901 cells into the right back limb of nude mice. On day 25, the average tumor volume in SGC7901-PrPC group was 1.86 ± 0.29 cm3, compared with 0.84 ± 0.15 cm3 of the control group and 1.04 ± 0.21 cm3 of the vector group (Fig. 1D )." provenance.