Primary Cell Culture of Hepatic Tumorous Cells From Routine Fine-needle Aspiration

• Sponsor: Kaohsiung Medical University Chung-Ho Memorial Hospital (Other)
• Overall Status: Completed
• CT.gov ID: NCT01549275
• Collaborator: Department of Health, Executive Yuan, R.O.C. (Taiwan) (Other)
• Enrollment: 105
• Location: 1
• Duration (Months): 39
• Patients Per Site Per Month: 2.7

Study Details

Study Description

Brief Summary

The purposes of this prospective study were to evaluate the successful rate of primary culture of hepatocellular carcinoma cells and cancer-associated fibroblasts from the residual specimens in routine fine-needle aspiration of hepatic tumor and the potential application of this method as an additional tool for personalized treatment of hepatocellular carcinoma patients.

Condition or Disease	Intervention/Treatment	Phase
Hepatocellular Carcinoma

Detailed Description

Fine-needle aspiration (FNA) cytology or biopsy of hepatic tumor is frequently applied in the collection of specimens because it is considered safe, efficacious, accurate and cost effective. Our previous study from small number of patients showed that specimens obtained from FNA had potential to be applied for primary culture of cancer cells. This study was to modified our method for primary culture of both hepatocellular carcinoma cells and cancer-associated fibroblasts in patients with tumor measuring larger than or equal to 3cm. All patients received one session of aspiration for the diagnosis of hepatic tumor. The aspirated specimens were applied for both cytologic and pathologic examinations at first. After then, 10 mL 0.9% NaCl was aspirated by the same needle connected with the same syringe and injected into 15 mL sterilized centrifugation tube. If the tube contained visible specimens, these specimens were sent for primary culture.

Study Design

• Study Type: Observational
• Actual Enrollment: 105 participants
• Observational Model: Case-Only
• Time Perspective: Prospective
• Official Title: Kaohsiung Medical University Hospital
• Study Start Date: Apr 1, 2010
• Actual Primary Completion Date: Jan 1, 2013
• Actual Study Completion Date: Jul 1, 2013

Outcome Measures

Primary Outcome Measures

1. Correlation Between the Growth Speeds of the Cultured Cells and the AJCC TNM Stage (7th Eds) at Entering of the Study. [28 days after plating of cells]

   Patients were divided into AJCC TNM staging < = IIIA and > = IIIB two groups. The incidences of patients with rapidly proliferative cultured cells in these two groups were compared. Rapidly proliferative group was defined as (1) growth area of cultured cells at the 28th day of primary culture exceeded two times of the growth area measured at the 14th day, or (2) growth area of cultured cells at the 28th day reached > 70% growth area of the flask. Based on the results from special stain, patients in rapidly proliferative group were further divided into patients with rapid proliferation of HCC cells alone, rapid proliferation of HCC cells with concomitant cancer-associated fibroblasts (CAFs) (HCC + CAFs) and CAFs alone.

Secondary Outcome Measures

1. Correlation Between the Growth Speeds of Cultured Cells and Worsening of AJCC TNM Stages or HCC Related Death 6 Months After Plating of Cells [6 months after plating of cells]

   104 Patients with complete follow-up data were further divided into receiving (1) curative treatment of HCC including operative resection and local ablation therapy, (2) palliative transcatheter arterial chemoembolization (TACE), and (3) supportive treatment.

Eligibility Criteria

Criteria

• Ages Eligible for Study: 20 Years to 90 Years
• Sexes Eligible for Study: All
• Accepts Healthy Volunteers: No

Inclusion Criteria:

• patients had residual aspirated specimens for primary culture.

Exclusion Criteria:

• patients did not have residual aspirated specimens for primary culture.

Contacts and Locations

Locations

	Site	City	State	Country	Postal Code
1	Kaohsiung Medical University Hospital	Kaohsiung		Taiwan	807

Sponsors and Collaborators

• Kaohsiung Medical University Chung-Ho Memorial Hospital
• Department of Health, Executive Yuan, R.O.C. (Taiwan)

Investigators

• Principal Investigator: Zu Y Lin, MD, Kaohsiung Medical University

Study Documents (Full-Text)

None provided.

More Information

Publications

• Lin ZY, Chuang WL, Chuang YH, Yu ML, Hsieh MY, Wang LY, Tsai JF. Discordant influence of amphotericin B on epirubicin cytotoxicity in primary hepatic malignant cells collected by a new primary culture technique. J Gastroenterol Hepatol. 2006 Feb;21(2):398-405.

• Responsible Party: Kaohsiung Medical University Chung-Ho Memorial Hospital
• ClinicalTrials.gov Identifier: NCT01549275
• Other Study ID Numbers: 990043
• First Posted: Mar 9, 2012
• Last Update Posted: Oct 8, 2015
• Last Verified: Jul 1, 2013

• Additional relevant MeSH terms: Carcinoma, Hepatocellular

Study Results

Participant Flow

Recruitment Details	From April 2010 to January 2013 in Cancer Center and Division of Hepatobiliary Medicine, Department of Internal Medicine, Kaohsiung Medical University Hospital
Pre-assignment Detail	A total of 114 patients signed the inform consents. However, 9 patients were excluded due to negative cytological and pathologic results of malignancy in 4 patients, adenocarcinoma in 2 patients and fungal contamination during the preparation of the specimens in 3 patients.

Arm/Group Title	AJCC TNM Staging < = IIIA HCC Patients	AJCC TNM Staging > = IIIB HCC Patients
Arm/Group Description	Tumor stage is based on the AJCC (American Joint Committee on Cancer) TNM staging system (2010, 7th edition). 76 patients with JACC TNM staging < = IIIA.	29 patients belonged to AJCC TNM staging > = IIIB.
Period Title: Overall Study
STARTED	76	29
COMPLETED	75	29
NOT COMPLETED	1	0

Baseline Characteristics

Arm/Group Title	AJCC TNM Staging < = IIIA HCC Patients	AJCC TNM Staging > = IIIB HCC Patients	Total
Arm/Group Description	Tumor stage is based on the AJCC (American Joint Committee on Cancer) TNM staging system (2010, 7th edition)	Tumor stage is based on the AJCC (American Joint Committee on Cancer) TNM staging system (2010, 7th edition)	Total of all reporting groups
Overall Participants	76	29	105
Age (Count of Participants)
<=18 years	0 0%	0 0%	0 0%
Between 18 and 65 years	31 40.8%	18 62.1%	49 46.7%
>=65 years	45 59.2%	11 37.9%	56 53.3%
Age (years) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [years]	66.4 (12)	59.6 (14)	63.1 (12.7)
Sex: Female, Male (Count of Participants)
Female	14 18.4%	5 17.2%	19 18.1%
Male	62 81.6%	24 82.8%	86 81.9%

Outcome Measures

1. Primary Outcome

Title	Correlation Between the Growth Speeds of the Cultured Cells and the AJCC TNM Stage (7th Eds) at Entering of the Study.
Description	Patients were divided into AJCC TNM staging < = IIIA and > = IIIB two groups. The incidences of patients with rapidly proliferative cultured cells in these two groups were compared. Rapidly proliferative group was defined as (1) growth area of cultured cells at the 28th day of primary culture exceeded two times of the growth area measured at the 14th day, or (2) growth area of cultured cells at the 28th day reached > 70% growth area of the flask. Based on the results from special stain, patients in rapidly proliferative group were further divided into patients with rapid proliferation of HCC cells alone, rapid proliferation of HCC cells with concomitant cancer-associated fibroblasts (CAFs) (HCC + CAFs) and CAFs alone.
Time Frame	28 days after plating of cells

Outcome Measure Data

Analysis Population Description

Comparison the incidence of patients with rapidly proliferative cultured cells between two groups

Arm/Group Title	AJCC TNM Staging > = IIIB	AJCC TNM Staging < = IIIA HCC Patients
Arm/Group Description
Measure Participants	29	76
Rapidly proliferative cultured cells	21 27.6%	29 100%
Rapid Proliferation of HCC with or without CAFs	14 18.4%	9 31%
Rapid Proliferation of CAFs alone	7 9.2%	20 69%

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	AJCC TNM Staging > = IIIB, AJCC TNM Staging < = IIIA HCC Patients
	Comments	Null hypothesis: There is no significant difference in the incidence of patients with rapidly proliferative cultured cells between two groups.
	Type of Statistical Test	Superiority or Other
	Comments
Statistical Test of Hypothesis	p-Value	< 0.005
	Comments
	Method	Chi-squared
	Comments	degrees of freedom =1

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	AJCC TNM Staging > = IIIB, AJCC TNM Staging < = IIIA HCC Patients
	Comments	Null hypothesis: There is no significant difference in the incidence of patients with rapidly proliferative cultured HCC cells with or without concomitant cancer-associated fibroblasts between two groups.
	Type of Statistical Test	Superiority or Other
	Comments
Statistical Test of Hypothesis	p-Value	< 0.005
	Comments
	Method	Chi-squared
	Comments	degrees of freedom = 1

Statistical Analysis 3

Statistical Analysis Overview	Comparison Group Selection	AJCC TNM Staging > = IIIB, AJCC TNM Staging < = IIIA HCC Patients
	Comments	Null hypothesis: There is no significant difference in the incidence of patients with rapidly proliferative cultured cancer-associated fibroblasts alone between two groups.
	Type of Statistical Test	Superiority or Other
	Comments
Statistical Test of Hypothesis	p-Value	> 0.1
	Comments
	Method	Chi-squared
	Comments	degrees of freedom = 1

2. Secondary Outcome

Title	Correlation Between the Growth Speeds of Cultured Cells and Worsening of AJCC TNM Stages or HCC Related Death 6 Months After Plating of Cells
Description	104 Patients with complete follow-up data were further divided into receiving (1) curative treatment of HCC including operative resection and local ablation therapy, (2) palliative transcatheter arterial chemoembolization (TACE), and (3) supportive treatment.
Time Frame	6 months after plating of cells

Outcome Measure Data

Analysis Population Description

Correlation between the growth speeds of cultured cells and worsening of AJCC TNM stages or HCC related death 6 months after plating of cells

Arm/Group Title	TNM Staging < = IIIA Patients Receiving Curative Treatment	TNM Staging < = IIIA Patients Receiving TACE	TNM Staging < = IIIA Patients Receiving Supportive Treatment	TNM Staging > = IIIB Patients Receiving TACE	TNM Staging > = IIIB Patients Receiving Supportive Treatment
Arm/Group Description	All patient belonged to Child A classification and BCLC (Barcelona Clinic Liver Cancer classification) degree A (14 patients) or B (8 patients).	All patients belonged to Child A classification and BCLC(Barcelona Clinic Liver Cancer classification) degree A or B.	5 patients belonged to BCLC (Barcelona Clinic Liver Cancer classification) degree B and the other 4 patients belonged to BCLC degree C or D.	All patients belonged to Child A classification and BCLC (Barcelona Clinic Liver Cancer classification) degree C.	All patients belonged to BCLC (Barcelona Clinic Liver Cancer classification) degree C or D.
Measure Participants	22	44	9	13	16
Rapidly proliferative cultured cells	10 13.2%	15 51.7%	3 2.9%	9 NaN	12 NaN
Rapid growth and worsening stage or death	0 0%	5 17.2%	3 2.9%	4 NaN	11 NaN
Slow growth and worsening staging or death	0 0%	2 6.9%	1 1%	0 NaN	2 NaN
Rapid growth of HCC cells and worsening	0 0%	2 6.9%	2 1.9%	4 NaN	8 NaN

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	AJCC TNM Staging > = IIIB
	Comments	null hypothesis: There is no significant difference in the incidence of change in AJCC TNM stages and survivals 6 months after plating of cells between patients with rapidly and slowly proliferative cultured cells.
	Type of Statistical Test	Superiority or Other
	Comments
Statistical Test of Hypothesis	p-Value	1
	Comments
	Method	Fisher Exact
	Comments	degrees of freedom = 1

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	AJCC TNM Staging < = IIIA HCC Patients
	Comments	Null hypothesis: There is no significant difference in the incidence of change in AJCC TNM stages and survivals 6 months after plating of cells between patients with rapidly and slowly proliferative cultured cells.
	Type of Statistical Test	Superiority or Other
	Comments
Statistical Test of Hypothesis	p-Value	0.032
	Comments
	Method	Fisher Exact
	Comments	degrees of freedom = 1

Statistical Analysis 3

Statistical Analysis Overview	Comparison Group Selection	TNM Staging < = IIIA Patients Receiving Supportive Treatment
	Comments	Null hypothesis: There is no significant difference in the incidence of change in AJCC TNM stages and survivals 6 months after plating of cells between patients with rapidly and slowly proliferative cultured cells.
	Type of Statistical Test	Superiority or Other
	Comments
Statistical Test of Hypothesis	p-Value	0.048
	Comments
	Method	Fisher Exact
	Comments	degrees of freedom = 1

Statistical Analysis 4

Statistical Analysis Overview	Comparison Group Selection	TNM Staging > = IIIB Patients Receiving TACE
	Comments	Null hypothesis: There is no significant difference in the incidence of change in AJCC TNM stages and survivals 6 months after plating of cells between patients with rapidly and slowly proliferative cultured cells.
	Type of Statistical Test	Superiority or Other
	Comments
Statistical Test of Hypothesis	p-Value	0.176
	Comments
	Method	Fisher Exact
	Comments	degrees of freedom = 1

Statistical Analysis 5

Statistical Analysis Overview	Comparison Group Selection	TNM Staging > = IIIB Patients Receiving TACE
	Comments	Null hypothesis: There is no significant difference in the incidence of change in AJCC TNM stages and survivals 6 months after plating of cells between patients with rapidly and slowly proliferative HCC cultured cells.
	Type of Statistical Test	Superiority or Other
	Comments
Statistical Test of Hypothesis	p-Value	0.021
	Comments
	Method	Fisher Exact
	Comments	degrees of freedom = 1

Statistical Analysis 6

Statistical Analysis Overview	Comparison Group Selection	TNM Staging > = IIIB Patients Receiving Supportive Treatment
	Comments	Null hypothesis: There is no significant difference in the incidence of change in AJCC TNM stages and survivals 6 months after plating of cells between patients with rapidly and slowly proliferative cultured cells.
	Type of Statistical Test	Superiority or Other
	Comments
Statistical Test of Hypothesis	p-Value	0.129
	Comments
	Method	Fisher Exact
	Comments	degrees of freedom = 1

Adverse Events

Time Frame	This study was carried out from April 2010 to January 2013.
Adverse Event Reporting Description	This study was to collect the residual specimens obtained from routine aspiration of hepatic tumor. This process did not have any influence on the participants. Serious and other Adverse Events were not collected/assessed."
Arm/Group Title	HCC Patients Underwent FNA of Tumor
Arm/Group Description	Patients who had residual specimens obtained from ultrasound-guided fine-needle aspiration of hepatic tumor measuring equal or larger than 3cm were included. The residual specimens were applied for primary culture and no additional aspiration of the tumor was performed solely for the collection of specimens for culture. Serious and other Adverse Events were not collected/assessed.
All Cause Mortality
	HCC Patients Underwent FNA of Tumor
	Affected / at Risk (%)	# Events
Total	/ (NaN)
Serious Adverse Events
	HCC Patients Underwent FNA of Tumor
	Affected / at Risk (%)	# Events
Total	0/0 (NaN)
Other (Not Including Serious) Adverse Events
	HCC Patients Underwent FNA of Tumor
	Affected / at Risk (%)	# Events
Total	0/0 (NaN)

Limitations/Caveats

[Not Specified]

More Information

Certain Agreements

Principal Investigators are NOT employed by the organization sponsoring the study.

There is NOT an agreement between Principal Investigators and the Sponsor (or its agents) that restricts the PI's rights to discuss or publish trial results after the trial is completed.