Expression of lola^GS88A8. psq^GS88A8 and Dl^UAS.cDa under the control of Scer\GAL4^ey.PH leads to eye tumor and metastasis.

The triple expression of Dl^UAS.cDa, lola^GS88A8 and psq^GS88A8 under the control of Scer\GAL4^ey.PH induces excessively enlarged eyes and eye tumors and macroscopically visible metastases, as compared to controls. The eye disc shows a disorganized epithelium with defects in the pattern of differentiated cell; there is a decrease in cell divisions (BrdU labelling and pH3 immunostaining) and an increase in apoptosis (Dcp1a immunostaining), as compared to controls.

Co-expression of Dl^Scer\UAS.cDa with lola and psq (using P{GSV1}lola^GS88A8, which is comprised of the lola^GS88A8 and psq^GS88A8 alleles) under the control of Scer\GAL4^ey.PU induces tumor-like overgrowths in the eye.

The tumor-like overgrowth phenotype seen when Dl^Scer\UAS.cDa is co-expressed P{GSV1}lola^GS88A8 (comprised of the lola^GS88A8 and psq^GS88A8) under the control of Scer\GAL4^ey.PU is seen with increased frequency when one copy of mirr^DFM3 is also present.

The tumor-like overgrowth phenotype seen when Dl^Scer\UAS.cDa is co-expressed P{GSV1}lola^GS88A8 (comprised of the lola^GS88A8 and psq^GS88A8) under the control of Scer\GAL4^ey.PU is seen with increased frequency upon co-expression of mirr^GD16318.

Co-expression of Dl^Scer\UAS.cDa, lola^GS88A8 and psq^GS88A8 using Scer\GAL4^ey.PH (the so-called 'eyeful' background) results in a delay in pupariation of 26.6 +/- 7.5 hours. This is reverted to a delay of 5.9 +/- 7.1 hours in a Ilp8^MI00727 background.

72.5% of flies expressing Dl^Scer\UAS.cDa, psq^GS88A8 and lola^GS88A8 under the control of Scer\GAL4^ey.PU have eye tumours, with 4.9% having macroscopically visible secondary tumour growths derived from the developing retina. The incidence of primary and secondary tumours in these "eyeful" flies is slightly enhanced by co-expression of ct^{dsRNA.cGa.Scer\UAS}, but is greatly enhanced if the "eyeful" flies also co-express both ct^{dsRNA.cGa.Scer\UAS} and BacA\p35^Scer\UAS.cUa.

Flies expressing Dl^Scer\UAS.cDa under the control of Scer\GAL4^ey.PH and also carrying the P{GSV1}lola^GS88A8 insertion (which causes overexpression of psq and lola) show an eye tumour phenotype. In addition, distant metastasis in the thorax and abdomen is seen in 15% of cases. Co-expression of mir-8^SC1 significantly reduces the primary eye tumour size, completely inhibits the incidence of metastases and results in a high incidence of adult lethality.

Scer\GAL4^ey.PH-mediated expression of Dl^Scer\UAS.cDa, psq^GS88A8 and lola^GS88A8 causes mild overgrowth of the eye (the 'eyeful' phenotype) at 18[o]C.

Scer\GAL4^ey.PH-mediated co-expression of Dl^Scer\UAS.cDa, psq^GS88A8 and lola^GS88A8 ("eyeful" flies) results in excessively enlarged eyes, and eye tumours occur in 57% of eyes, with 3% of flies showing macroscopically visible metastases derived from the developing retina.

Co-expression of ato^Scer\UAS.cJa almost completely suppresses the formation of eye tumours seen in 'eyeful' (Scer\GAL4^ey.PH, Dl^Scer\UAS.cDa, psq^GS88A8, lola^GS88A8) flies.

Co-expression of ato^{dsRNA.Scer\UAS.J} results in a dramatic increase in tumour and metastases incidence in 'eyeful' (Scer\GAL4^ey.PH, Dl^Scer\UAS.cDa, psq^GS88A8, lola^GS88A8) flies.

Co-expression of ato^{Scer\UAS.T:Rep-en.T:Hsap\MYC} results in a dramatic increase in tumour and metastases incidence in 'eyeful' (Scer\GAL4^ey.PH, Dl^Scer\UAS.cDa, psq^GS88A8, lola^GS88A8) flies: differentiated eye tissue is lost in 30% of the eye fields, and tumors are seen in 100% of the remaining eyes. These tumors include large patches of undifferentiated tissue.

Co-expression of ato^Scer\UAS.cJa in 'eyeful' third instar eye discs (Scer\GAL4^ey.PH, Dl^Scer\UAS.cDa, psq^GS88A8, lola^GS88A8) results in dramatically increased apoptosis, as measured by decay staining.

Scer\GAL4^ey.PH-mediated co-expression of Dl^Scer\UAS.cDa, psq^GS88A8 and lola^GS88A8 ("eyeful") third instar eye discs exhibit increased cell proliferation (as measured by phospho-histoneH3) posterior to the second mitotic wave.

Co-expression of ato^Scer\UAS.cJa reduces ectopic proliferation in "eyeful" (Scer\GAL4^ey.PH Dl^Scer\UAS.cDa, psq^GS88A8 and lola^GS88A8) third instar eye discs.

Co-expression of ato^{Scer\UAS.T:Rep-en.T:Hsap\MYC} increases ectopic proliferation in "eyeful" (Scer\GAL4^ey.PH Dl^Scer\UAS.cDa, psq^GS88A8 and lola^GS88A8) third instar eye discs.

Co-expression of ato^{Scer\UAS.T:Rep-en.T:Hsap\MYC} in a Scer\GAL4^ey.PH Dl^Scer\UAS.cDa background leads to the appearance of significant numbers of proliferating cells in the posterior region of third instar eye discs.

"Eyeful" (Scer\GAL4^ey.PH Dl^Scer\UAS.cDa, psq^GS88A8 and lola^GS88A8) third instar eye discs exhibit disorganization of the epithelium.

Co-expression of ato^Scer\UAS.cJa restores the size and organization of "eyeful" (Scer\GAL4^ey.PH Dl^Scer\UAS.cDa, psq^GS88A8 and lola^GS88A8) third instar eye discs.

Co-expression of ato^{Scer\UAS.T:Rep-en.T:Hsap\MYC} severely worsens the retinal patterning of "eyeful" (Scer\GAL4^ey.PH Dl^Scer\UAS.cDa, psq^GS88A8 and lola^GS88A8) third instar eye discs, to the extent that the morphogenetic furrow is difficult to discriminate.

Co-expression of dap^Scer\UAS.cUa leads to a significant inhibition of the tumor occurrence and a partial reduction in the metastasis seen in 'eyeful' (Scer\GAL4^ey.PH, Dl^Scer\UAS.cDa, psq^GS88A8, lola^GS88A8) flies.

Co-expression of bsk^DN.Scer\UAS in the 'eyeful' (Scer\GAL4^ey.PH, Dl^Scer\UAS.cDa, psq^GS88A8, lola^GS88A8) background leads to tumors in 61% of eyes and an approximately 7-fold increase in metastasis.

Co-expression of Jra^Scer\UAS.cEa reduces tumor and metastasis incidence in the 'eyeful' (Scer\GAL4^ey.PH, Dl^Scer\UAS.cDa, psq^GS88A8, lola^GS88A8) background.

Co-expression of bsk^DN.Scer\UAS in a ato^Scer\UAS.cJa 'eyeful' (Scer\GAL4^ey.PH, Dl^Scer\UAS.cDa, psq^GS88A8, lola^GS88A8) background leads to a suppression of the inhibitory effects of ato^Scer\UAS.cJa on the 'eyeful' tumor phenotype, restoring tumor formation and enhancing metastasis.

Flies expressing Dl^Scer\UAS.cDa under the control of Scer\GAL4^ey.PH and also carrying the P{GSV1}lola^GS88A8 insertion (which results in the lola^GS88A8 and psq^GS88A8 alleles being expressed under the control of Scer\GAL4^ey.PH) usually develop tumours in the eyes. Additionally, in approximately 30% of the flies, secondary eye growths are seen throughout the body, with the whole body filling up with eye tissue in some cases. The secondary eye growths have ragged borders, indicating invasion of the mutant tissue into the surrounding normal tissue. Eye discs show massive uncontrolled overgrowth in animals expressing Dl^Scer\UAS.cDa under the control of Scer\GAL4^ey.PH and also carrying the P{GSV1}lola^GS88A8 insertion (which results in the lola^GS88A8 and psq^GS88A8 alleles being expressed under the control of Scer\GAL4^ey.PH). In most of the discs, the epithelial cells have lost their apical-basal polarity and some have a disrupted basement membrane and grow without differentiating. Wing discs of animals expressing Dl^Scer\UAS.cDa under the control of Scer\GAL4^dpp.blk1 and also carrying the P{GSV1}lola^GS88A8 insertion (which results in the lola^GS88A8 and psq^GS88A8 alleles being expressed under the control of Scer\GAL4^dpp.blk1) show massive overgrowth and mutant cells are found invading adjacent regions of the disc. Wing discs of animals expressing Dl^Scer\UAS.cDa under the control of Scer\GAL4^Bx-MS1096 and also carrying the P{GSV1}lola^GS88A8 insertion (which results in the lola^GS88A8 and psq^GS88A8 alleles being expressed under the control of Scer\GAL4^Bx-MS1096) show massive overgrowth, which initiates in the dorsal region but progressively extends to occupy the whole disc. Flies expressing Dl^Scer\UAS.cDa under the control of Scer\GAL4^ey.PH and also carrying psq^GS88A8 and lola^rev6 have reduced eye tumour size (compared to flies expressing Dl^Scer\UAS.cDa under the control of Scer\GAL4^ey.PH and also carrying psq^GS88A8 and lola^GS88A8) but show sporadic secondary growth of eye tissue. The eye tumour phenotype of flies expressing Dl^Scer\UAS.cDa under the control of Scer\GAL4^ey.PH and also carrying the P{GSV1}lola^GS88A8 insertion (which results in the lola^GS88A8 and psq^GS88A8 alleles being expressed under the control of Scer\GAL4^ey.PH) is dominantly suppressed by one of Rpd3⁰⁴⁵⁵⁶, Su(var)3-9¹⁷ or E(z)⁷³¹, is dominantly enhanced by Rbf¹⁴, but is not enhanced by trx^unspecified or ash1^unspecified. The eye tumour phenotype of flies expressing Dl^Scer\UAS.cDa under the control of Scer\GAL4^ey.PH and also carrying the P{GSV1}lola^GS88A8 insertion (which results in the lola^GS88A8 and psq^GS88A8 alleles being expressed under the control of Scer\GAL4^ey.PH) is suppressed by co-expression of Rbf^Scer\UAS.cDa.