Next we wanted to understand the phenotypic changes associated with the in vitro transformed vimentin expressing cells. BP treated vimentin positive cells showed a significant increase in its in vitro invasive potential, although no significant difference was seen in the migration and proliferation potential of the two clones (Fig. 3A-E). Exogenously expressed vimentin levels remained unchanged till the end of BP treatment (Supplementary Fig. S4A and B). Surprisingly, neither BP treated vimentin expressing nor its vector control cells showed endogenous induction of vimentin expression, which suggests that perhaps it occurs very late in the process of transformation and further treatment with BP for several weeks, might be required to achieve this stage. The E-cadherin expression remained downregulated while expression of Oct-4, Sox2 and Nanog showed significant increase in BP treated vimentin positive cells (Fig. 3F, Supplementary Fig. S5A). Additionally, BP treated vimentin positive cells showed no difference in the levels of p53, Ras, ?-catenin and EMT regulators (Snail, Slug, Twist), while a decline was seen in the protein level of involucrin (Supplementary Fig. S5A-C). Interestingly, tumorigenic potential of BP treated vimentin positive cells was significantly higher, as reflected by the increase in volume of subcutaneous tumors, in NOD-SCID mice. On the other hand control cells failed to form tumors and in one case they formed a cyst (Fig. 3G and Supplementary Table S5). Furthermore, IHC analysis confirmed the differential levels of vimentin between the vimentin positive and its control group (Fig. 3H).
3.4 Significant correlation was seen between high vimentin-low E-cadherin expression and lymph node metastasis, in OSCC patients
Significant correlation was seen between expression levels of vimentin-E-cadherin and demographic/clinical parameters like age (p = 0.008), stage (p = 0.001), node status (p