Antiangiogenic Effects and Therapeutic Targets of Azadirachta indica Leaf Extract in Endothelial Cells

Table 1

(a) Upregulation and (b) downregulation in the RNA expression levels of 30 genes in HUVECs treated with 20.0 and 40.0 μg/mL of EENL for 24 hours, validated by real-time PCR. The endogenous GAPDH mRNA levels were measured as internal controls. Mean ± SD of the fold change in the expression levels of genes compared with the respective vehicle controls were shown. Assay IDs of the genes used for validation were from Applied Biosystems. Accession number and major molecular function for each gene are also shown.

(a)

Gene	Fold upregulation (20 μg/mL)	Fold upregulation (40 μg/mL)	Assay ID	Function
AKR1B10	17.68±3.25	48.31±8.68	Hs00252524_m1	Aldo-keto reductase activity
AKR1C1	6.70±1.51	23.13±5.43	Hs00413886_m1	Oxidoredutase activity
AKR1C2	2.95±0.55	23.31±5.07	Hs00912742_m1	Oxidoredutase activity
AKR1C3	2.73±0.87	4.47±2.67	Hs00366267_m1	Oxidoredutase activity
ALDH3A2	2.01±1.33	3.97±2.96	Hs00166066_m1	Aldehyde dehydrogenase
ALOX5	7.74±2.50	1.97±1.42	Hs01095330_m1	Lipoxygenase activity
ATF3	86.64±12.44	277.09±16.31	Hs00231069_m1	Transcription factor
CHAC1	54.21±4.2	156.95±14.98	Hs00225520_m1	Protein binding
CLU	2.53±1.04	4.47±2.44	Hs00156548_m1	Protein binding
CSTA	4.58±0.42	9.71±2.05	Hs00193257_m1	Protease binding
DDIT3	27.43±8.54	203.64±20.65	Hs00358796_g1	Nucleic acid binding
DMRT1	6.68±1.45	21.91±3.11	Hs00232766_m1	Transcription factor
EGF	10.27±4.08	3.52±1.80	Hs01099999_m1	Signal transducer
EGR1	4.06±1.67	87.38±15.36	Hs00152928_m1	Transcription factor
FOXC1	1.96±1.24	5.91±3.68	Hs00559473_s1	Transcription factor
GCLM	3.48±1.76	15.00±9.08	Hs00157694_m1	Glutamate-cysteine ligase
GPNMB	4.75±0.53	38.43±6.90	Hs01095669_m1	Integrin binding
HMOX1	11.01±2.30	26.19±5.21	Hs01110251_m1	Heme oxygenase activity
ID2	4.48±1.09	19.82±4.24	Hs00747379_m1	Transcription repressor
JDP2	5.56±2.14	24.51±3.22	Hs00185689_m1	Transcription factor
LAMP3	6.08±2.82	2.35±1.09	Hs00180880_m1	Integral to membrane
LY96	2.14±0.23	3.82±2.42	Hs00209771_m1	Receptor activity
PEG3	21.38±8.36	87.45±30.33	Hs00377844_m1	Transcription factor
S100P	102.62±31.88	570.45±45.47	Hs00195584_m1	Calcium-dependent protein
SESN2	7.34±4.98	23.62±6.63	Hs00230241_m1	Cell cycle arrest
SPINK1	247.34±23.27	465.02±53.98	Hs00162154_m1	Endopeptidase inhibitor
SPRR1A	4.22±4.49	70.57±20.12	Hs00954595_s1	Structural molecule
TRIM16	2.08±1.19	4.32±2.56	Hs00414879_m1	DNA binding
TUBA1A	2.06±1.27	3.46±0.80	Hs00362387_m1	Structural molecule
WDR19	2.80±1.32	9.43±4.26	Hs00228414_m1	Transmembrane signaling

(b)

Gene	Fold downregulation 20 μg/mL	Fold downregulation 40 μg/mL	Assay ID	Function
ANKRD12	3.83±1.84	21.63±6.80	Hs00209530_m1	Ion channel inhibitor
ASPM	2.36±0.63	5.69±2.34	Hs00996458_m1	Calmodulin binding
CDC2	5.50±2.22	9.39±4.12	Hs00938777_m1	Protein binding
CDC25A	15.65±3.26	2.09±1.78	Hs00947994_m1	Phosphatase activity
CDCA4	7.98±2.45	3.34±1.85	Hs00743989_s1	Cell division
CENPE	78.27±25.90	52.49±16.37	Hs00156507_m1	Chromatin binding
CHEK1	24.20±3.23	60.16±29.80	Hs00176236_m1	Protein Kinase
COL12A1	3.62±0.59	2.64±1.23	Hs00329355_s1	Structural molecule
DLGAP5	12.34±5.89	4.78±2.63	Hs00207323_m1	Phosphatase activity
DPP4	15.39±4.21	15.78±2.62	Hs00175218_m1	Aminopeptidase
DTL	3.88±1.40	1.55±0.80	Hs00978565_m1	Protein binding
E2F8	75.55±30.92	80.34±19.30	Hs00226635_m1	Transcription factor
EBP	9.75±3.21	3.26±1.05	Hs00198130_m1	Sterol isomerase
FBXO5	4.25±2.17	3.75±0.54	Hs00559989_m1	Protein binding
FOLH1	5.92±2.39	5.23±1.31	Hs00379515_m1	Carboxypeptidase
HIST1H4C	8.34±5.41	2.73±0.73	Hs00543883_s1	DNA binding
HSP90B1	26.21±12.03	30.88±13.74	Hs00427665_g1	Nucleotide binding
ITGAV	13.50±2.31	2.68±1.86	Hs00233790_m1	Receptor
KIF14	1.93±1.24	15.28±4.89	Hs00208408_m1	Microtubule motor
MAD2L1	5.55±3.25	7.83±2.16	Hs01554515_g1	Protein binding
METAP2	5.04±1.20	15.45±5.21	Hs01127366_m1	Aminopeptidase
NRIP1	24.88±6.20	45.47±14.54	Hs00942766_s1	Transcription coactivator
POLA1	15.62±3.22	4.39±2.87	Hs00213524_m1	Nucleotide binding
PRIM2	11.83±5.41	2.32±0.33	Hs00386277_m1	DNA primase
RRM1	5.87±1.72	11.23±4.21	Hs01040698_m1	Ribonucleoside diphosphate reductase
SKP2	17.52±11.90	16.43±4.23	Hs00180634_m1	Ubiquitin-protein ligase
TOP2A	15.53±3.89	10.34±2.11	Hs01032127_g1	Topoisomerase activity
TOP2B	2.03±1.48	2.25±0.79	Hs00172259_m1	Topoisomerase activity
TPR	1.35±1.91	4.66±1.30	Hs00162918_m1	Protein kinase
UHRF1	10.73±2.84	3.41±1.52	Hs00273589_m1	Transcription factor