# Estrogen Induces c-myc Transcription by Binding to Upstream ERE Element in Promoter
## 3 Results
### Age Related c-myc Regulation in Breast Cancer Lymphocytes  
The relative expression of c-myc in patients was normalized against healthy control. Patients were divided in four distinct age groups (average age at onset of the disease = \(\pm\)46.6 years): age group 21-35 (Figure 4A), age group 36-45 (Figure 4B), age group 46-50 (Figure 4C), and age group > 60 (Figure 4D). We observed that overall, the mean RE of c-myc is relatively high in patients as compared to healthy control, as shown in Figure 4E (\(p\)-value \(<0.0001\)). Research analysis provided a clearer view of elevated c-myc RE in an age-related fashion. We observed that after certain age, i.e., age > 60, the c-myc-related expression is less elevated as compared to age groups < 60 years, which could be due to the decreased level of estrogen. We calculated the relative risk from binomial pairwise comparison after performing the Pearson chi-square goodness of fit test, and results showed that the relative risk for age group 36-45 and age group 46-60 was >1, while the relative risk for age groups 21-35 and >60 was <0.143. This is due to the age-specific expression pattern of estrogen. Estradiol level were also significantly elevated in patients with \(p\) value < 0.001 as compared to age matched healthy control (Figure 5).  
Figure 3: Correlation studies between c-myc, estradiol, and BMI status of breast cancer patients Using Psych package in R software. (**A**): Positive correlation (R = 0.46) exists in c-myc and BMI status. (**B**): Positive correlation (R = 0.50) between estradiol and BMI status (image depicting R\({}^{2}\) value is then converted to R correlation coefficient. \({}^{*}\) Correlation is significant at 0.01 level (two tailed).  
Figure 4: **(A–E)** Comparison of mean Relative Expression (RE = 2\({}^{\Delta\Delta\text{CI}}\)) of c-myc in breast cancer patients and healthy controls (RE calculated using Livak method from two-fold Ct values). Error bars indicate standard deviation. Asterisks indicate a significant difference determined using [PERSON]’s \(t\)-test (* \(p>0.05\)). **(A)** RE of age group 21 to 35 years patients \(n=13\), SEM = 4.1, (Average 2\({}^{\Delta\Delta\text{CI}}\) = 10.32) and healthy control (Average 2\({}^{\Delta\Delta\text{CI}}\) = 0.99). \(n=23\), SD = 0.1, Stq of sample size = 4.8, SEM = 0.01, df = 1, \(p\)-value = 0.0027. **(B)** 2\({}^{\Delta\Delta\text{CI}}\) of age group 36 to 45 years. patients (Average 2\({}^{\Delta\Delta\text{CI}}\) = 5.81) and healthy control (Average 2\({}^{\Delta\Delta\text{CI}}\) = 1.95). \(n=39\), SD = 3.6, Stq of sample size = 6.2, SEM = 1.5, df = 1, \(p\)-value = 0.01. **(C)** 2\({}^{\Delta\Delta\text{CI}}\) of age group 46 to 60 years patients (Average RE = 7.2) and healthy control (Average RE = 0.36). \(n=41\), SD = 0.48, Stq of sample size = 3.46, SEM = 0.14, df = 1, \(p\)-value < 0.0001. **(D)** RE of age group > 60 years. patients (Average RE = 2.49) and healthy control (Average RE = 0.8). \(n=25\), SD = 1.76, Stq of sample size = 5, SEM = 0.07, df = 1, \(p\)-value = 0.0134. **(E)** Overall RE of patients (Average RE = 8.4 \(n=116\), SD = 14.9, SEM = 0.25) and healthy control (Average RE = 0.99, \(n=76\), SD = 2.17) df = 1, \(p\)-value < 0.0001 (** = \(p\)-value 0.001, *** = \(p\)-value 0.0001).