The effect of andrographolide on Human papillomavirus type 16 (HPV16) positive cervical cancer cells (SiHa)

Abstract

Background: Infection with a high-risk human papillomavirus (HR-HPV), especially, HPV16 is the most important risk factor for cervical cancer. Oncoproteins (E6 and E7) of HR-HPV play an important role in cervical cancer development involved by several mechanisms including degradation of tumor suppressor protein p53 and anti-apoptosis. Andrographolide is a diterpenoid lactone isolated from a traditional herbal medicine,Andrographis paniculata. It has been reported to induce apoptosis in different cancer cell lines. However, the effects on regulation of HPV16 transcription, oncogene expression and induction of cervical cancer cells apoptosis are still unclear. This study investigated the effects of andrographolide on HPV16 transcription activity, E6 oncogene expression and p53 protein level as a downstream process involved in cell apoptosis. Methods: Sub-cytotoxicity of the andrographolide compound was determined by MTT assay. C33A cell line, an HPV-negative cervical cancer cell line, was transfected by reporting vector containing long control region of HPV16 European and Asian variant and treated with compound for analysis of promoter transcription. SiHa cell line, an HPV16-positive cervical cancer cell line, was treated with the compound for 24 and 48 hr. The oncogene E6 and p53 expression were analyzed by SYBR Green real-time PCR and western blot. Cell apoptosis was analyzed by flow cytometry. Results: Andrographolide exhibited cytotoxic effects with IC50 values of 152.34 M and 142.23 M, whereas sub-cytotoxic values of 9.71 M and 13.88 M in C33A and SiHa cells, respectively. The result demonstrated that sub-cytotoxic concentration of andrographolide suppressed LCR transcription activity of both HPV16 European and Asian variant in transfected-C33A cells. Andrographolide also significantly inhibited E6 oncogene expression in SiHa cells at 24 h post treatment. Importantly, inhibitory effect was increased by increasing concentration, but not time increasing. After 48 h post treatment, the p53 was restored expression in SiHa cells. In addition, andrographolide also significantly induced apoptosis of SiHa cells in concentration-dependent manner. Conclusion: This result demonstrated that the andrographolide induced SiHa cells apoptosis via suppression of HPV16 transcription activity, leading to decreased E6 oncoprotein and restored p53. These findings imply that the andrographolide may be an effective agent for cervical cancer prevention and treatment.

Andrographis paniculata (Burm. f.) Wall. ex Nees: An Updated Review of Phytochemistry, Antimicrobial Pharmacology, and Clinical Safety and Efficacy

“…A recent study also reported a similar number of unique secondary metabolites [169]. Among these compounds, about 80 compounds showed different types of pharmacological activities, including antibacterial [109,139,[170][171][172][173][174][175], anti-biofilm [88,176], antiviral (i.e., chikungunya virus [91], Anti-HIV [177][178][179], anti-influenza [99,180], Herpes simplex virus 1 [83,99,181], dengue virus serotype 1 [92][93][94]182], anti-Epstein Barr virus [97], pestivirus and flavivirus [90], human papillomavirus type 16 [96]), anti-fungal [109,183], antiparasitic (i.e., antimalarial [95,184,185], anti-Leishmaniasis [98]), antiproliferative [85][86][87]120,122,124,127,128,177,186], cytotoxicity [84][85][86]…”

“…[39,176,192,193]. [139,[170][171][172][173], anti-biofilm [88,139], anti-CHIKV [91], Anti-HIV [177][178][179], anti-influenza [99,180], anti-HSV-1 [83,99,181], anti-DVS-1 [92][93][94]182], anti-EBV [97], anti-HPV-16 [96], anti-HBV [186], anti-HCV [194], antimalarial [95,184,185], anti-Leishmaniasis [98] [190,191] AeP…”

Andrographis paniculata(Burm. f.) Wall. ex Nees: A Review of Ethnobotany, Phytochemistry, and Pharmacology

“…Researchers investigated significant antiviral activity of AP besides other pharmacological activities in last two decades. Although they reported antiviral activity against limited viruses, such as dengue virus serotype 1 (DENV-1) [ 201 ], human papilloma virus type 16 (HPV16) [ 202 ], herpes simplex virus type 1 (HSV-1) [ 203 ], influenza A virus [ 102 ], and HIV [ 53 , 55 , 80 , 204 ], their findings were very encouraging and noteworthy considering the life threatening role of these viruses in human community. The hot aqueous aerial parts extract of AP was reported for its significant antiviral activity to reduce the percentage of HIV antigen-positive H9 cells [ 55 ].…”

“…Recently, Tang et al [ 201 ] reported that the methanol extract of AP possesses significant inhibition activity against DENV-1 in vitro assay. Another study has revealed that andrographolide suppressed HPV16 transcription activity, leading to the reduction of E6 oncoprotein and restored p53 [ 202 ].…”

Andrographolide Inhibits Lytic Reactivation of Epstein-Barr Virus by Modulating Transcription Factors in Gastric Cancer

“…f.) Nees (family Acanthaceae) contains diverse compounds such as labdane diterpenoid lactones, flavonoids, and miscellaneous compounds and is widely used as a traditional medicine in Asian countries [5,7]. Anti-viral activity of andrographolide has been shown for a number of viruses including herpes simplex virus [8], dengue virus (DENV) [9], chikungunya virus (CHIKV) [10], human papillomavirus (HPV) [11], hepatitis B virus (HBV) [12], and influenza A [13].…”